Haloethisterone compounds



3,100,204 HALOETHISTERONE COMPOUNDS Arthur E. Oberster, Canton, Ohio, Roger E. Beyler, Carbondale, 111., and Lewis H. Sarett, Princeton, N.J., assignors to Merck & Co., Inc., Rahway, N.J., a corporation of New Jersey No Drawing. Filed Apr. 20, 1960, Ser. No. 23,393 5 Claims. (Cl. 260-239.55)

This invention is concerned generally with novel steroids and processes of preparing the same. More particularly, it relates to novel 2l-haloethisterones, 2l-halonorethisterones, and closely related compounds which possess useful therapeutic properties as orally and parenterally active progestational agents. This invention also relates to pharmaceutical compositions containing these novel steroid compounds.

In accordance With the present invention, the novel 21- haloethisterones (17a-haloethynyl-4-androstene-175-01-3- ones) and the 21-halonorethisterones (l7a-haloethynyll9-nor-l7fi-ol-3-ones) are prepared from a 17a-ethynyl- 3-oxygenated-androstene-17 8-01 by first protecting the oxygen functions present at the C-3 and 017 positions of the steroid molecule with suitable protecting groups, and then halogenating to form the l7a-haloethyny1 derivative. The protecting groups at C-3 and C-17 are then removed.

This process may be schematically represented as follows, starting with a 17m-ethynyl-4-androstene-17 9-01-3- one in which R is methyl, or the corresponding l9-norsteroid in which R is hydrogen:

HO CECE Step1 0 HO CECE II-A 0 0 CECH Step2 O atent O 3,100,204 Patented Aug. 6, 1963 Step3 O III HO ozox H0 0502:

IV-A IV-B no 'CECX rv-o When starting with the -19-nor-17oz-ethynyl-4-androstene-17fl-ol-3-one, in which R is hydrogen, Compound H-A (and likewise Compounds 11-8 and III) are mixtures of the A and the A -androstenes, and in the case of the A500) compound, the hydrogen (R) shown at 0-10 is present at (3-4; Removal of the protecting groups in Step 3, using concentrated HCl gives only the 3-ketoA androstene (IV-A), but using approximately aque HO 'CECH If 'thestartin'g material is the l7ez -ethynyl-- androstene- 3;3,17.B-diol (1'), the product (IV which is obtained-on removal of the protecting groups is the 17mhaloethynyl -5-androstene-3 8,17,8-diol. This compound may be converted into the '17a-ha1oethynyl-4-androstene- 17B-ol-3-o'ne 2l-haloethisterone) by oxidation, for example, with aluminum isopropoxide and cyclohexanone (or acetone) 'inbenzene. The choice of the 3-oxygenatedsteroid used as starting material is based on practical cons'id'eration, availability and the like.

In the first step of our process the oxygen functions present atthe C-3 and (3-17 positions'of the steroid molecule are protected. In apreferred embodiment of our invention a 3-keto group is blocked by forming the 3- ethylenedioxy-derivative by reaction with excess ethylene glycol in benzene, toluene or ethylene dichloride solvent in the presence of acid catalysts suchas p-toluenesulfonic or sulfuric acid, the water by-product being continuously removed.

The 3-keto group may also be converted into the 3- ethylenedioxy derivative by exchange dioxolanation, which involves acid-catalyzed transfer of the ethylene glycol portion of simple 2,2-dialkyl-1,3-dioxolanes, such as 2,2'-d-irnethyl-1,3-dioxolane (acetone ethylene ketal) or,

4 better, 2-methyl-2-ethyl-l,3-dioxolane (butanone ethylene ket-al), on reaction of the ketones either in an inert sol- ,vent, as benzene, or simply in excess reagent.

Other cyclic ketal derivatives can be used in our process for protecting a ketone group at C-3. In general, we have found that the lower alkylenedioxy derivatives wherein the hydrocarbon group contains not more than seven carbon atoms, such as the ethylenedioxy, t-rimethylenedioxy, propylenedioxy and butylenedioxy derivatives are most suitable. However, in place of using a lower alkylenedioxy substituent to block or protect the keto substituent, we can use other derivatives readily hydrolyzable to keto, such as an enol ether monothioketal, or a dithioketal derivative for this purpose.

The 17,8-01 group, and the 313,17I3-dio1 groups in the steroid molecule are protected by reaction with dihydropyran in the presence of an acidic reagent, such as p-toluenesul fonyl chloride, to form the corresponding tetrahydropyranyl ether. This reaction takes place, for example, on addition of p-toluenesulfonyl chloride to a solution of the steroid in an excess of dihydrcpynan. The reaction mixture is stirred at room temperature for 16 to 64 hours. The product may be recovered by neutralizing the reaction mixture with dilute base, and then extracting with ether. The ether extracts are washed with Water, dried and evaporated to dryness. The crude product is dissolved in a solvent such as petroleum ether, and chromatographed over alumina to give the corresponding tetrahydropy-nanyl derivative.

In Step 2 of our invention, the steroid starting material, protected at the 3- and the 17-positions as indicated above, is treated with a suitable halogenating agent. In the preferred embodiment of our invention, the steroid compound, dissolved in a tertiary alcohol, is first treated with a potassium alcoholate of that tertiary alcohol, toform the 21-potassium derivative of the steroid compound, and the latter compound is then halogenated.

To form the 21-chloro-derivative, the steroid is dissolved in tbutyl alcohol and treated with potassium-tbutoxide and then with t-butyl-hpyochlorite.

To form the 2l-b-romo-derivative, the steroid is dissolved in -t-butyl alcohol and treated with potassium t-butoxide and then with N-bromosuccini-mide. The choice of the tertiary \alcohol reagents is determined by practical considerations, such as availability, solubility and the like.

The 2l-halo-rsteroid compounds are conveniently recovering by adding water to the reaction mixture, and extracting with ether. The ether extracts are then washed with water, dried, and evaporated to dryness in vacuo. The residual material thus obtained is separated by chromatography on alumina to afford the Zl-halo-steroid.

iIn Step 3 of our process, the protecting groups at C-3 and C-l7 are removed, preferably by treatment with acid. Removal of the protecting groups using concentrated HCl gives the corresponding 3-keto-A -androstene. To carry out this reaction, a solution of the steroid in methanol is stirred with concentrated HCl for approximately one hour at room temperature. The methanol may then be removed under reduced pressure. The steroid is conveniently recovered by ether extraction. The ether extract is dried, evaporated to dryness and the residual material crystallized from a suitable solvent.

When star-ting with the l9-nor-17a-ethynyl-4-androstene-l7l3-ol-3-one, in which R is hydrogen, removal of the protecting groups from the mixture of the l9-nor-A and A -androstenes present in III by treatment with approximately 70% aqueous acetic .acid for about one hour at room temperature-gives a mixture of the S-keto- 19-norA the 3-keto-l9-nor-A and the 3-keto-l9- nor-A androstenes. These compounds can 'be separated by chromatography over acid-washed alumina and elution with mixtures of ether and petroleum ether.

The l7owhaloethynyl-5-androstene-3,3,l7fi-diol (IV') may be converted into the 17a-hal0ethynyl-4-androstene 17,8-ol-3-one (ZI-halothisterone) (V') by Oxidation with aluminum isopropoxide and cyclohexanone in benzene. This oxidation is carried out by dissolving the steroid in a solvent such as a mixture of cyclohexanone and benzene and reacting in an inert atmosphere with a solution of aluminum isopropoxide in benzene at 80-90" C. for 2-16 hours. To recover the product, the solution is cooled, a few drops of water are added, and the resulting aluminum hydroxide is filtered off. The filtrate is concentrated on the steam bath under reduced pressure, and the residual material is crystallized from a suitable solvent.

In accordance with the present invention, the novel 21-haloethisterones are prepared from the 17a-ethynyl-5- androstene-3/5',17fl-diol by first reacting the latter compound with dihydropyran in the presence of p-toluenesulfonyl chloride to give the 17u-ethynyl-5-androstene-- 3B,17,8adiol-bis-tetrahydropyranyl ether.

The 170: ethynyl 5 androstene 318,176 diol bistetrahydropyranyl ether is chlorinated by dissolving in t-butyl alcohol and treating with a solution of potassium t-butoxide and then with -t-buty-l hypochlorite to give the 170: ch-loroethynyl 5 androstene 3,8,175 diol bistetrahydropyranyl ether. The latter compound is then hydrolyzed with concentrated HCl to give 17oc-Chl0l0- ethynyl-5-androstene-3B,17B-diol, which is then oxidized with aluminum isopropoxide and cyclohexanone in benzene to give the 17oa-chloroethynyl-4-androstene-17,8-01-3- one (2l-chloroethisterone) The 17 a ethynyl 5 iandrostene 318,175 diol bistetrahydropyranyl ether is brominated by dissolving the steroid in t-butyl alcohol and treating with a solution of potassium t-butoxide and then With N-bromosuccinimide to give the 17a-bromoethyny1-5-androstene-3[3,17fi-di01- bis-tetrahydropyranyl ether. The latter compound is then hydrolyzed with concentrated I-ICl to give 17a-bromoethynyl-5-androstene-3B,17 3-diol, which is then oxidized with aluminum isopropoxide and cyclohex-anone in benzene to give the 17a-bromoethynyl-4-androstene-17,8-01- 3-one (21-bromo-ethisterone).

The novel 2l-halonorethisterones are prepared from norethisterone (17a ethynyl 19 nor 4 androstene- 17fl-ol-3-one) by reaction first with ethylene glycol in the presence of p-toluenesulfonic acid to .give a mixture of the A and the A -17a-ethynyl-3-ethylenedioxy-19-norandrostene-17fl-ol, and then with dihydropyran to give the corresponding tetrahydropyranyl ether.

The mixture of the A and the A -17u-ethynyl3- ethylenedioxy 19 nor androstene 175 01 tetrahydropyranyl ether is chlorinated by dissolving the steroid in t-butyl alcohol and treating with a solution of potassium t-butoxide and then with t-butylhypoohlorite to give a mixture of the A and the A -17a-ch'loroethyny1-l9- nor androstene 17B o1 3 one tetrahydropyranyl ether, which mixture is then hydrolyzed with acid. Hydrolysis of the mixture with concentrated I-ICl gives the 170: chloroethynyl 19 nor 4 androstene 17,8 o1 3- one (2l-chloronorethisterone). Hydrolysis of the mixture with 70% aqueous acetic acid gives a mixture of the A the A and the A -17a-chloroethynyl-19-nor-androstene-17/3-ol-3-ones, which mixtur is separated by chromatography.

The mixture of the A and the A -17a-ethyny1-3- ethylenedioxy-19-nor-androstene-17B-oltetnahydropyra nyl ether is brominated by dissolving the steroid in tbutyl alcohol and treating with a solution of potassium tbutoxide and then with N-bromosuccinimide to give a mixture of the A and the A -17a-bromoethyny1- 19-nor-androstene-17,8-0l-3-one tetrahydropyranyl ether. The latter compound is then hydrolyzed with acid. Hydrolysis of the mixture with concentrated HCl gives the 17a-bromoethynyl-19-nor-4-androstene-17B-ol-3-one (ZI-bromonorethisterone). Hydrolysis of the mixture with 70% aqueous acetic acid gives a mixture of the A, the A500) and the A -17a-bromoethynyl-19-nor-andro- 6 stene-17B-ol-3-ones, which mixture is separated by chromatography.

In addition to the above described 17a-haloethynyl-4- androstene-17fi-ol-3-ones, this invention contemplates the preparation of certain derivatives thereof, and in particular the novel steroid compounds which may be chemically represented as follows:

wherein the 1,2-positions and the 6,7-positions of the steroid molecule may be double bonded or saturated, and wherein R is hydrogen, a methyl group R is a lower alkyl or alkanoyl group, X is CECCI, CECBI, CH CHCL CH CHBr, CH CH Br or CH CH Cl, and Y is methyl, chloro or fluoro.

This invention also contemplates the preparation of the 17/8-lower alkyl esters and ethers of the Not-haloethynyl-19-nor-5 (10) -androstene-17,8-01-3-ones, as well as the products which are formed on hydrogenating the 17a-haloethynyl group of the aforesaid compounds to the 17a-CH=CHX, and to the 17ocCH CH X groups, where X represents chlorine or bromine.

The 17a-haloethynyl group of the ZI-haloethisterones and the 21-halonorethisterones (VI) is reduced to the corresponding chlorovinyl derivative (VI) on hydrogenation with a Lindlar catalyst, and to the corresponding chloroethyl derivative (VII) on hydrogenation using a heavy metal catalyst such as PtO VIII The 6a-methyl-2l-h'aloethisterone is prepared by reacting the 17a-haloethynyl-5-androstene-3fl,17fl-dio1 with monoperphthalic acid to give the 17a-hal0ethynylandrostatic-3B,l7B-diol-5,6-oxide, which on reaction with methyl magnesium iodide yields 6,8-methyl-l7m-lraloethynyl-andI'OStaHe-3I3,5a,l7fl-l1'l0l. The latter compound is oxidized with an oxidizing reagent {prepared from chromium trioxide and sulfuric acid to give 6 B-methyl-17a-haloethynyl-androstane-Sa,l7p-diol-3-one, which on treatment with aqueous sodium hydroxide gives the 6a-methy1-17ahaloethynyl-4-andros-tene-17fl-ol-3-one.

The 6a-methyl-17a-haloethynyl-4-androstene-17 3-01-3- one is dehydrogenated at C-l by means of selenium dioxide, or alternately by microbiological methods, to give the (Six-methyl-l7a-haloethynyl4,4-androstadiene-17Bwl- 3-one.

The Got-methyl-17a-haloethynyl-4-androstene-176-01-3- one is reacted with chloranil to give 6-methyl-17a-haloethynyl-4,6-androstadiene-l7p-ol-3-one, which compound is dehydrogenated at 01, by means of selenium dioxide, or alternately by microbiological methods, to give 6- methyl 17cc haloethynyl 1,4,6 androstatriene 17 p ol-3-one.

The 17/3 lwer alkyl ethers of the 17B-hydroxy-17ahaloethynyl-3-keto-steroids are prepared by the reaction of the corresponding 17fl-hydroxy-steroid with a lower alkyl halide and silver oxide in a solvent such as dimethylformam-ide. The lower alkyl halides which may be used for this purpose includes methyl iodide, ethyl iodide, n-propyl iodide, n-butyl iodide and the like.

The l7B-lower alkanoyl ethers of the 17B-hydroxy-17ahaloethynyl-3-keto-steroids are prepared by the reaction of the corresponding 17fi-hydroxy-steroid with a lower alkanoic acid anhydride in the presence of an organic base such as pyridine. The lower acid anhydrides which may be used for this purpose include acetic anhydride, propionic anhydride, butyric anhydride, and the like.

The 170: haloethynyl 1,4 :androstadiene 17,8 ol 3 one acetate is prepared by reacting 17u-haloethynyl-4-andostane-17B 0l-3-one with acetic anhydride in pyridine to give the corresponding acetate, and then converting the latter compound into the 17a-haloethynyl-L4-androstadiene-17fl-o1-3-one acetate by means of selenium dioxide, or alternately by microbiological methods.

The 17u-haloethynyl-1,4-androstadiene-l7fl-methoxy-3- one is prepared by reacting l7u-haloethynylt-androstene- 17fl-ol-3-one with methyl iodide and silver oxide, using dimethylformamide as solvent, to give the l7ot-haloethynyl-4-androstene-17B-methoxy-3-0ne and then converting the latter compound into the 17-a-haloethynyl-L4-androstadiene-17fi-rnethoxy-3-one by means of selenium dioxide, or alternately by microbiological methods.

The Goa-chloro-17a-haloethynyl-l,4-andro stadiene-17B- ol-3-one acetate is prepared by reacting 17a-haloethyny1-4- androstene-l7B-ol-3-one with acetic anhydride and ptolnenseulfonic acid to give the l7m-haloethynyl-3,5-androstadiene-3,l7t3-diol-diacetate. The latter compound is reacted with N-chlorosuccinimide to give the 6a-chlorol7u-haloethynyl-4-androstene-17fi-ol-3-one acetate, which is converted into the 6ot-chloro-17u-haloethynyl-l,4-androstadiene-l7fi ol-3-one acetate by reaction with selenium dioxide, or alternately by microbiological methods.

The GQ-ChtlOI'O-170L-113l06il1YI1Yl-1,4-aI1d1'OStadlIl$-17[3- methoxy-3-one is prepared by reacting l7a-haloethynyl-4- landrostene-l7fi-ol-3-one with methyl iodide and silver oxide, using dimethylformamide as solvent, to give the 170: haloethynyl 4 andros-tene 176 methoxy 3 one. The latter compound is then converted into the 17a-haloethynyl-3,S-androstadiene-l7fl-methoxy-3-ol acetate, by reaction with acetic acid and p-toluenesulfonic acid. The acetate so formed is treated with N-chlorosuccinimide to give the 6a-chloro-l7oz-haloethynyl-4-androstene-175-methoxy-3-one, which compound is dehydrogenated by selenium dioxide, or alternately by microbiological methods, to the Got-chloro-l7a-haloethynyl-L4- androstadiene-l7B-methoxy-3-one.

The fi'chloro-17a-haloethynyl-4,o-androstadiene-175-01- 3-one acetate is prepared by reaction of l7a-haloethynyl- 4-androstene-l7B-ol-3-one with chlonanil to give the h-aloethynyl-4,6-androstadiene-l7B-ol-3 one (A -21-haloethisterone), which is converted into the l7a-haloethynyl- 4androstene-17 8-ol-3-one-6J-oxide on reaction with perbenzoic acid. The latter compound is reacted with HCl in chloroform solution to give the 6-chloro-17m-haloethynyl-4,6-androstadiene-l7fi ol-3-one which forms the acetate on treatment with acetic anhydride in pyridine, and the corresponding 17 fl-methoxy-derivative on reaction with methyl iodide and silver oxide. The 6-chloro-l7ahaloethynyl-4,6-androstadiene-l7B ol-3-one acetate is dehydrogenated to the 6-chloro-l7u-h-al-oethynyl-1,4,6-androstatriene-17B-ol-3-one acetate on reaction with selenium dioxide, or alternately by microbiological methods.

The 6a-fiuoro-l7a-haloethynyl-4-androstene-l7,8=ol-3- one acetate is prepared from the 17 a-h-al-oethynyl-3,5-androstadiene-3,l7fi-diol diacetate by reaction with perchloryl fluoride followed by acid treatment. The 6afiuoro-l7a-haloethynyl-4-androstene-l7fi-ol-3-one acetate is dehydrogenated at 0-1 by means of selenium dioxide, or alternately by microbiological methods, to give 60:- fluoro-l7a haloethyny-l 1,4 androstadiene-l7 3-ol-3-one acetate.

The 6a-fluoro 17oz haloethynyl-4-androstene-176-01- 3-one acetate is reacted with chloranil to give 6-flu'orol7rx-haloethynyl-4,6 androstadiene-l7 8-ol-3-one acetate, and the latter compound is dehydrognated at C-1 by means of selenium dioxide, or alternately in microbiological methods, -to give 6-fluoro-17u-haloethynyl-l,4,6-androstatriene-l7fi-0l-3-one acetate.

The l7a-haloethynyl 3,5 androstadiene-17fi-methoxy- 35-01 acetate is reacted with perchloryl fluoride followed by acid treatment to give Goa-fluoro-l7a-haloethynyl-4-androstene-17fitmethoxy-3-one, which compound may be dehydrogenated at O1, by means of selenium dioxide, or alternately by microbiological methods, to give 6a-fluorol7a-ha-loethynyl-1,4-androstadiene- 1 7t? -meth oxy-3 -one.

The 60c fiuoro 17a haloethynyl-4-androstene-175- methoxy-3-one is converted into 6-fiuoro-l7a-haloethynyl- 4,6- andr0stadiene-l7fl-rnethoxy-3-one by the use of chlor-anil. The 6-fluoro 17a haloethy-nyl-4,6-androstadiene- 17fl-methoxy-3-one is dehydrogenated at (2-1 by means of selenium dioxide, or alternately by microbiological methods, to yield 6-fiuoro-l7a-haloethynyl-1,4,6-androstatriene- 17 fi-methoxy-B -one.

The 17 a-haloethynyl 4 andro'stene-l7dol-3-one is reduced to 21-halo-4,2O-pregnadiene-17,8-01-3-one by hydrogenation in ethyl acetate, using a Lind-lar catalyst, and to 21-halo-4-pregnene-17,8-01-3-one by hydrogenation in ethy-l alcohol using a platinum oxide catalyst.

The 6a-methyl-derivatives of the 2l-halonoreth-isterones are prepared by first converting the 2l-halo-norethisterones (l7oc-h aloethynyl-19-nor-4-androstene-17-ol-3-one) into the l7a-haloethy1nyl-l9 nor-5-androstene-3B,17,6-dioll7fl-acetate by reacting the l7a-haloethynyl-l9-nor-4-androstene-l7 8-ol-3-one with acetic anhydride and p-toluenesulfonic acid to give the l7u-haloethynyl-l9 nor-3,5- androstad-iene-3,l7fi-diol-diacetate and then reacting the latter compound with sodium borohydride. Oxidation of 1'7u-haloethynyl 19' nor-S-androstene 3fl,l'7fl-diol-l76- acetate withmonopenphthalic acid yields l7m-haloethynyl- 19-nor-androstane 35,175 diol 5,6-oxide-17fl-acetate. The later compound is reacted with methyl magnesium iodide to form op-methyl-lh-haloethynyl-l9-nor-androstane-3B,5a,17 8-triol, Which on oxidation with chrornic oxide in acetone gives -6;8-methyl-l7zx-haloethynyl-19-norandrostane-Su,17fl-dio1-3-one. The latter compound is treated with sodium hydroxide to form 6a-me-thyl-l7ehaloethynyl-l9-nor-4-androstene-17/3-ol-3-one.

The 6a-methyl-l7a-hal-oethy-nyl 19 nor-4-androstenel7 8-ol-3-one' is converted to 6rx-methyl-l7a-haloethynyll9-nor-4-androstene-17.;i-methoxy-3-one by reaction with methyl iodide and silver oxide. The 6u-rnethyl-l7a-haloethynyl-4-androstene-l7fi-ol-3-0ne is treated with acetic 9 anhydride in pyridine to give 6a-methyl-17-haloethynyl- 19-nor-4-androstene-17,B-ol-3-one acetate.

The 6ct-chloro-17a-haloethynyl-19-nor 4 androstene- 17/3-01-3-one acetate is prepared from the 17a-haloethynyl- 19-nor-3,5-androstadiene-3,l7fi-diol acetate by reacting with N-chlorosuccinimide.

The Ge-fluoro-17a-haloethynyl 19 nor 4 androstene-l7p-ol-3-one acetate is prepared from the 17a-haloethynyl-l9-nor 3,5 androstadiene-3,17fi-diol diacetate by reacting with perchloryl fluoride followed by acid treatment.

The 6oc-ChlOTO-17oc-h3l06thYHYl 19 nor 4 androstene-17/8-methoxy-3-one is prepared from the 17oc-' aloethynyl l9 nor 4 androstene-17B-ol-3-one by reacting first with silver oxide and methyl iodide to form the 170:- haloethynyl-l9-nor-4-androstene 17f methoxy-3-one. The latter compound is then reacted with acetic anhydride and p-toluenesulfonic acid to give the l7u-haloethynyl- 19-nor-3,5-androstadiene-l7p methoxy 3 o1 acetate, which is converted into the 6a-chloro-17u-haloethynyl-19 nor-4-androstene-17fl-methoxy-3 one on treatment with N-chlorosuccinimide.

The 6a-fluoro-l7a-haloethynyl 19' nor 4 androstene-175-methoxy-3-one is obtained by reacting the 17ahaloethynyl-l9-nor-3,5-androstadiene-l7p methoxy 3-01 acetate with perchloryl fluoride followed by acid treatment.

The l7a-haloethynyl-19-nor-4 androstene 175-01-3- one is reduced to 21-halo-19-nor-4,20-pregnadiene-17B-ol 3-one by hydrogenation in ethyl acetate, using a Lindlar catalyst and to 21-halo-l9-nor-4-pregnene-l7fl ol-3-one by hydrogenation in ethyl alcohol, using a platinum oxide catalyst.

The 17a-haloethynyl-19-nor-5(10)-androstene 17 3-01- 3-one forms the acetate on treatment with acetic anhydride in pyridine, and the corresponding 17 B-methoxy-derivative on reaction with methyl iodide and silver oxide. The 17a-haloethynyl-19-nor (10) androstene-17B-ol- 3-one is reduced to the corresponding chlorovinyl derivative (the 21-halo-19-nor-5(10),20-pregnadiene-17B-ol-3- one) on hydrogenation with a Lindlar catalyst, and to the corresponding chloroethyl derivative (the 21-halo-19-nor- 5(l0)-pregnene-17[3-ol-3-one) on hydrogenation using a heavy metal catalyst such at P-tO A further embodiment of our invention comprises novel pharmaceutical compositions containing these 21- haloethisterones and 21-halonorethisterones.

Various changes and modifications may be made in the present invention, certain preferred embodiments of which are herein disclosed, without departing from the scope thereof; to the extent that these changes and modifications are Within the scope of the appended :claims, they are to be considered -a part of this invention.

Example 1 Twenty mg. of p-toluenesulfonyl chloride is added to 400 mg. of 17a-ethynyl-5-androstene-3[3,17p-diol in 20 ml. of dihydropyran. The resulting mixture is allowed to stand at room temperature overnight. A 2.5 N NaOH solution is added until the mixture is slightly alkaline. Water is then added and the aqueous phase extracted with 4 portions of ether, each containing approximately 50 ml. The combined ether layers are washed with water, dried over N21 SO and evaporated under reduced pressure to give about 725 mg. of a non-crystalline product. The product dissolved in petroleum ether is chromatographed on 60 g. of neutral alumina and the chromatogram eluted with a 7:3 mixture of petroleum ether; ether to give 400 mg. of crystalline product, the 170:- ethynyl-S-androstene 3 3,170 diol-bis-tetrahydropyranyl ether,

A solution of about 4 grams of 17e-ethynyl-5-androstene-3fi,17,B-diol bis-tetrahydropyranyl ether in 75 ml. of t-butyl alcohol is prepared. About 1.1 equivalents of a 10 1.0 molar potassium tbutoxide is added and the resulting mixture refluxed for one hour, with stirring, and then cooled. About 1.84 ml. of t-butyl hypochlorite is then added in one portion and the reaction mixture is left stirring at room temperature overnight. About ml. of water is added and the resulting aqueous mixture is extracted with four portions of ether, each containing approximately 200 ml. The combined layers are washed with water, dried Over sodium sulfite, filtered and evaporated to dryness in vacuo. The residual material is dissolved in petroleum ether and chromatographed on g. of alumina. Elution with petroleum ether gives about 3.10 grams (a 70% yield) of crystals of 17a-chloroethynyl-5-androstene-3B,17fl-diol-bis-tetrahydropyranyl ether. The crude product shows infrared pea-ks at 4.414 and 2.9a.

A solution of about 3 g. of the l7aachloroethynyl-5- androstene-3B,17,8-diol-bis+tetrahydropyranyl ether in ml. of methanol is prepared. To this solution is added 2.5 ml. of concentrated hydrochloric acid and the reaction mixture is stirred for about 1 hour at room temperature. The methanol is then removed by evaporation under reduced pressure until the product crystallizes. Approximately 100 ml. of water is then added and the resulting product is extracted with four portions of ether, each containing about 200 ml. The combined ether extract is washed with water, dried over sodium sulfate and evaporated to a crystalline residue. The residual crystalline material is recrystallized several times from ether to give about 1.58 g. of 17aachloroethynyl-5-androstene-3B,17,8-diol which has the following properties: M.P. 195 C. Analysis.-Calculated for C H O Cl: C, 72.30; H, 8.38; Cl. 10.16. Found: C, 71.64; H, 8.63; Cl. 10.48.

One hundred mg. of 17e chloroethynyl-5-androstene- 35,175-dio1 is dissolved in 1.0 ml. of cyclohexanone and 10 ml. of benzene in a flask fitted with a magnetic stirrer and a reflux condenser. About 5 ml. of the benzene is distilled and a stream of dry nitrogen is passed through the system and maintained throughout the reaction time. Then 0.5 ml. of a 10% solution of aluminum isopropoxide in benzene is added and the reaction mixture is maintained at reflux temperature for 4 hours. The solution is cooled, 5 drops of water are added and the resultant aluminum hydroxide is filtered off. The filtrate is taken to dryness under reduced pressure. The material is dissolved in ether, filtered, and the filtrate concentrated to give about 37 mg. of crude 17a-chloroethyny1-4-androstene--01-3-one, M.P. 178183 C. Recrystallization from ether gives about 25 mg. of the purified product, M.P. 182-184" C. Chromatography of all mother liquors on 3 g. of alumina and elution of the chromatogram with ether gives an additional 20 mg. of product, M.P. 181-184 C. Total yield 45 mg. The product has the following properties:

U.V. A352 241 m 6 15,000. LR. xiii 6.18y

Example 2 To a solution of 482 mg. of 17a-ethynyl-5-androstene- 3,8,17,8-diol-bis-tetrahydropyranyl ether in 10 ml. of tertiary-butyl alcohol, is added about 1.1 equivalents of a 1.0 molar potassium t-butoxide. The resulting mixture is refluxed for one hour, with stirring, and then cooled. 196 mg. of N bromosuccinimide is then added and the reaction mixture is stirred at room temperature for about 18 hours. The entire reaction mixture is dissolved in water and then extracted with 3 portions of ether, each containing approximately 50 ml. The combined ether extracts are washed with three portions of a saturated solution of NaHCO each portion containing approximately 25 mL, then with 3 portions of water, each containing about 25 ml. The ether layer is dried over sodium sulfate, filtered and evaporated to dryness. The oily residue is filtered through 20 g. of aluminum oxide to give 407 mg. of oily material which is dissolved in petroleum ether and chromatographed on 30 g. of acetone activated alkaline alumina. Elution with a 9:1 mixture of petroleum ether and ether yields 87 mg. of l7a-bromoethynyl-S-androstene 313,175 diol-bis-tetrahydropyranyl ether. An infrared spectrum of this material shows X 4.51;.

To' a solution of 17a bromoethynyl--androstene-3,8, l7fi-diol-bis-tetrahydropyranyl ether in 40 ml. of methanol is added 0.8 ml. of concentrated I-ICl,and the reaction mixture is stirred for one hour at room temperature. The methanol is then removed under reduced pressure. Water is added and the resulting solution is extracted with 3 portions of ether, each portion containing approximately 75 ml. The combined ether extracts are washed three times with approximately 50 ml. of water, dried over sodium sulfate, filtered and evaporated to dryness. The residual material is crystallized to give 230 mg. of 17a-bromo-ethynyl-S-androstone-3p,l7/3-diol which has thefollowing properties: MP. 214-215 C.

IR. A53 2.7, 2.89, 4.55 1.

Analysis-Calculated for C H O Br: C, 64.10; H, 7.43; Br, 20.32. Found: C, 62.40; H, 7.65; Br, 20.50.

17ot-bromoethynyl-5-androstene-3 8,17p-diol (195 mg.) is dissolved in 1.95 ml. of cyclohexanone and 20. ml. of benzene, using a flask fitted with a magnetic stirrer and a reflux condenser. After 3 ml. of benzene'is distilled, a stream of dry nitrogen is passed through the system, and maintained throughout the entire reaction time. After cooling to room temperature, there is added 0.98 ml. of a solution of aluminum isopropoxide in benzene, and the reaction mixture is refluxed for 3 hours and cooled to room temperature. Ten drops of water are added and the reaction mixture is filtered. The filtrate is taken to dryness. The residue is chromatognaphed on g. of acetone activated acid-washed alumina and 85 mg. of product is eluted with a mixture of seven parts ether to three parts petroleum ether. The 17a-brornoethynyl-4- androstene-l7B-ol-3-one so obtained has the following properties:

U.V. XQ TQ 2.40 11151, 6 15,700, LR. REE? 2.9, 4.51, 6.0, 6.211.

Analysis.-Calculated for C H O Br: C, 64.45; H, 6.95; Br, 20.42. Found: C, 65.16; H, 7.30; Br, 21.44.

Example 3 To a solution of one gram of l7u-ethynyl-l9-nor-4- androstene-l7fi-ol-3-one dissolved in 75 ml. of benzene is added 7.5 ml. of ethylene glycol, together with 50 mg. of p-toluenesulfonic acid. The reaction mixture is heated at reflux with a water separator for hours. The reaction mixture is cooled, and about 10 ml. of bicarbonate solution is added. The reaction mixture is extracted with 3 portions of ether, each portion containing about 100 ml. The combined extracts are Washed with water, dried over sodium sulfate and evaporated to dryness to give a mixture of the A and A -17a-ethynyl-3-ethylenedioxy-19-nor-androstenel 7 8-ols.

To a solution of the total crude material from the above reaction in 10 ml. of dihydropyran which has been distilled from sodium metal, is added with stirring, 120 mg. of p-toluenesulfonyl chloride, and the stirring is continued at room temperature for 64 hours. Saturated sodium bicarbonate solution (10 ml.) is then added to the reaction mixture. The mixture is then extracted with 3 portions of ether, each portion containing about 50 ml. The extracts are combined, washed with Water and dried over sodium sulfate, and evaporated to dryness in vacuo. The crude oil (about 3.5 g.) is dissolved in petroleum ether and chromatographed on 100 g. of acetone activated alkaline alumina. The chromatogram is eluted with mixtures of ether and petroleum ether to give 1.043 g. of a 12 mixture of the A and A -l7a-ethynyl-3-ethylenedioxy- 19-nor-androstene-l7fl-ol-tetrahydropyranyl ethers. The infrared spectrum shows bands at 2.9g and 9.0

To a stirred solution of 2.130 g. of the mixture of the A and A -17u-ethynyl-3-ethylenedioxy-l9-nor-androstene-l7fl-ol-tetrahydropyranyl ethers, dissolved in 50 ml. of dry t-butanol is added 5.5 ml. of 1 M potassium-t-butoxide. The reaction mixture is heated at reflux for ninety minutes and then cooled to room temperature. Then 0. ml. of t-butyl hypochlorite is added. The react-ion mixture is left to stir at room temperature for about 16 hours. About ml. of Water is added to the reaction mixture and the aqueous solution is extracted with three portions of ether, each portioncontaining about 200 ml. The extracts are combined, washed 'with water, dried over sodium sulfate, filtered and evaporated to dryness under reduced pressure. About 2. 2 g. of crude product is obtained. This is chromatographed on 100 g. of alkaline alumina. The material dissolved in petroleum ether to which a small amount of benzene has been added is chromatographed on alumina and eluted with etherpetroleum ether mixtures. 1.9 g. of a semi-crystalline product, the 17ot-chl0ro-ethynyl-3-ethy1enedioxy-l9-nor- 5 [and 5 (10)]-androstene-l7B-ol-tetrahydropyranyl ether, is obtained. infrared absorption shows bands at 4.45; and strong absorption in the 910;t region.

To a stirred solution of 1.9 grams of 17a-chloroethynyl- 3-ethylenedioxy-l9-nor-5 [and 5(10) ]-androstene-l7fi-oltetrahydropyranyl dissolved in 41 ml. of methanol is added 3 ml. of concentrated hydrochloric acid and 2.1 ml. of water. The reaction mixture is stirred at room temperature for 2 /2 hours. The methanol is then removed under reduced pressure. The residue is dissolved in about 200 ml. of ether and washed with water until the washings are neutral. The ether solution is dried over sodium sulfate and evaporated to dryness. The crystalline residue, recrystallized from a mixture of ether and methylene chloride, gives 1.2 g. of 17a-chloroethynyl-19-nor-4-androstene-17/8-ol-3-one, M.P. 194200 C. A sample recrystallized for analysis from ethyl acetate has a melting point of 198201 C. and shows the following properties:

U.V.t 1x32 240 III 1., 15,000, LRJ h2g2" 2.91.4, 4.48 1. 6.05 1, 6.2g

Analysis.-Calculated for C I-I O Cl: C, 72.20; H, 7.57; Cl, 10.65. Found: C, 72.27; H, 7.57; Cl, 9.90.

Ten mg. of the mixture of 17ct-chloroethynyl-3-ethylenedioxy-A [and A -androstene-l 7fl-ol-tetrahydropyranyl ether is treated with 1 cc. of 70% aqueous acetic acid for 1 hour at room temperature. The product is neutralized with aqueous sodium hydroxide and extracted with ether. The ether extract is dried and concentrated in vacuo. The crude product is chromatographed on acid-Washed alumina and eluted with mixtures of ether and petroleum ether to give the 17a-ethy-nyl-5 10) -androstene- 17fl-ol-3-one.

Example 4 3.2 ml. of potassium t-butoxide is added to 849 mg. of 17a-ethynyl-3-ethylenedioxy-l9-nor-5 [and 5 10) 1 -androstene-l7,8-ol-tetrahydropyranyl ether, prepared as in -Example 3, in 28 ml. of t-butyl-alcohol. This mixture is refluxed for 75 minutes and then cooled to room temperature. 1.136 g. of :N-bromosuccinimide is added and the reaction mixture is then stirred at room temperature for about 17 hours. The reaction mixture is added to about 60 ml. of water and extracted with 3 portions of ether, each portion containing about 75 ml. The ether extracts are combined, washed with 3, 50 ml. portions of saturated sodium bisulfite solution, and then with water until the washings are neutral. The other solution is dried over sodium sulfate, filtered and evaporated to dryness to yield the crude 17a-bromoethynyl-3-ethylenedioxy-l9-nor- 5[and 5 (10)]-androstene-l7fi-ol-tetrahydropynanyl ether which may be used in the next step without purification.

The crude steroid is dissolved in 25 ml. of methanol and 0.2 ml. of concentrated hydrochloric acid. The mixture is stirred at room temperature for 2 /2 hours. The methanol is then removed under reduced pressure, water is added, and the reaction mixture is extracted with 3, 75 ml. portions of ether. The ether layer is washed with water, dried over sodium sulfate, filtered and evaporated to dryness. This gives 737 mg. of crude oil which is chromatographed on 44 g. of acetone activated acidwashed alumina. Elution with a mixture of ether and petroleum ether gives a crystalline material which is recrystallized from ether to yield about 75 mg., M.'P. ISO-182 of 17a-bromoethynyl-19-nor-4-androstene-17flol-3-0ne which has the following properties:

U.V.: M12? 239 m Emol, 16,300, I.R.: A531? 6.1 and 6.2;].

Example 5 Ten grams of 17a-chloroethynyl-S-androstene-3B,17/3- diol in 150 ml. of tetrahydrofuran is treated with 100 ml. of 0.9 m. monoperphthalic acid in ethyl acetate and allowed to stand overnight at room temperature. The reaction mixture is diluted with ethyl acetate and washed sequentially with aqueous sodium bicarbonate, sodium bisuliite, and sodium bicarbonate solution. The organic layer is dried and concentrated in vacuo. The product is fractionally crystallized from aqueous ethanol to yield the 17a-chloroethynyl-androstane-3/5,17,B-diol5,6a-oxide.

Five grams of l7a-chloroethynyl-androstanc-3B,17 6- diol-5,6a-oxide in benzene is added to a reagent prepared from 3.4 g. of magnesium, 10 ml. of methyl iodide and 45 ml. of ether. The mixture is stirred and 70 ml. of solvent is removed by distillation. After refluxing for 5 hours, the mixture is cooled, acidified with dilute hydrochloric acid and the organic layer washed to neutrality. The organic phase is dried and concentrated in vacuo. Crystallization from aqueous ethanol gives the 6 8-methyl- 17a-chloroethynyl-androstane-Sfi,5a,17fi-triol.

The oxidizing reagent is prepared by diluting 2.7 g. of CrO and 2.3 ml. of concentrated sulfuric acid to 10 ml. with water. A solution of 1.90 g. of 6B-methyl-l7achloroethynyl-androstane-36,541,17fl-triol in 300 ml. of acetone is cooled to and treated with 1.85 ml. of the oxidizing reagent. The solution is diluted with ice water and extracted with ether. The ether extract is washed sequentially with water, and aqueous soduim bicarbonate and then dried and concentrated in vacuo, to give 65- methyl 17a chloroethynyl androstane a,17[3 diol- 3-one. The crude product is dissolved in 150 ml. of methanol, 75 ml. of 1 N sodium hydroxide is added and the mixture is stirred at room temperature under nitrogen for 24 hours. The mixture is concentrated to half volume under vacuo at 25 C. and then poured into ice water. The product is extracted with ether. The ether layer is washed with water, dried and concentrated. The concentrate is chromatographed on acid-washed alumina and the product eluted with ether-petroleum ether mixtures to yield 6a-rnethyl-17a-chloroethynyl-4-androstene-175-01- 3-one.

In accordance with the above procedures, but star-ting with the 17a-bromoethynyl-5-androstene-3;3,17B-diol in place of the 6a-methyl-l7a-chloroethynyl-4-androstene- 17/3-ol-3-one, there is obtained the 6a-methyl-17a-bromoethynyl-4-androstene-l7p3-ol-3-one.

Example 6 To mg. of GOL-IIIClhYl-t-Chl0IO6thYl1Yl-4-3Ildl0- stene-17fl-ol-3-one in 5 m1. of t-butanol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is washed successively with aqueous sodium bicarbonate, ammonium sulfide, dilute ammonia water, water, dilute hydrochloric acid and water, and then dried over magnesium sulfate. The extract is treated with activated charcoal and then concentrated to dryness. Crystallization of the residue from a mixture of acetone and ether gives 6a-methyl-17achloroethynyl-l,4-androstadiene-17/3-ol-3-one.

In accordance with the above procedures, but starting with the 6a-methyl-17tt-bromoethynyl-4- androstene-17pol-3-one in place of the 6ot-methyl-17a-chloroethynyl-4-an drostene-l7B-ol-3-one, there is obtained the 6a-methyl- 17m-bromoethynyl-1,4-androstadiene-17fl-ol-3-one.

Septomyxa afilnis (ATCC 6737) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Shefiield Farms), 5% glucose, and 0.5% corn steep liquor. After a 48 hour incubation at 28 C., 10 mg. of 6u-methyl-l7a-chloroethynyl-4-androstene-17/3-ol-3-one is added to each shake flask as a dimethylformamide solution (100 mg./ml.). After a conversion period of '24 hours, the cells are removed by rfiltration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The 6a-methyl-l7a-chloroethynyl-1g4-androstadiene- 17,8-o1-3-one is readily crystallized from the concentrate. Descending paper chromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

Bacillus sphaericus (ATCC 12488) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Sheflield Farms), 5% glucose, and 0.5% corn steep liquor. After a 24 hour incubation at 28 C., 10 mg. of 6ot-methyl-l7a chloroethynyl-4-androstene-l7[3-ol-3-one is added to each shake flask as a dimethylformamide solution (100 mg/ ml.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The 60c methyl 17c: chloroethynyl 1,4 androstadiene-l7fl-ol-3-one, is readily crystallized from the concentrate. Descending paper chromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A-dehydrogenation product.

In accordance with the above procedures, but using the 6ot-methyl 17oz brornoethynyl 4 androstene 17B- ol-3-one in place of the 6a-methyl-17u-chloroethynyl-4- androstene-l7B-ol-3-one, there is obtained the 6a-Inethyl- 17abromoethynyl-1,4-androstadiene-17fl-ol-3-one.

Example 7 A suspension of 1 g. of 6a-methyl-17-chloroethynyl-4- androstene-l7fi-ol-3-one and 2.4 g. of chloranil in 30 ml. of ethyl acetate and 6 ml. of acetic acid is stirred and refluxed for 16 hours. The reaction mixture is cooled and filtered. The product is washed sequentially with aqueous bisulfite, aqueous potassium hydroxide and water. The organic layer is dried and concentrated in vacuo. Chromatography yields 6-methyl-17a-chloroethynyl-4,'6- androstadiene-l7,8-ol-3-one.

Example 8 To 100 mg. of 6methyl-17a-chloroethynyl-4,6-androstadiene-17-5-ol-3-one in ml. of t-butanol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed for an additional 24 hours. The product is filtered and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is washed successively with aqueous sodium bicarbonate, ammonium sulfide, dilute ammonia water, water, dilute hydrochloric acid and water, and is then dried over magnesium su-lfate. The extract is treated with activated charcoal and then concentrated to dryness. Crystallization of the residue from a mixture of acetone and ether :gives -methyll'la-chloroethynyl l,4,6-androstatriene-17fi-ol-3-one.

In accordance with the above procedures, but starting with the 6-methyl-l7a-bromoethynyl-4,6-androstadienel76-ol-3-one in place of the 6-metl1yl-l7ot-chloroethynyl- 4,6-androstadiene-17B-ol-3-one there is obtained the 6- methyl 17oz bromoethynyl 1,4,6 androstatriene 17B- ol-3-one.

Septomyxa afiinis (ATCC 6737) is inoculated from a slant into 250 ml. sha-ke flasks containing 50 ml. of the following medium: 2% Edamin (Sheffield Farms), 5% glucose, and 0.5% corn steep liquor. After a 48 hour incubation at 28 C., mg. of 6-methyl-l7a-chloroethynyl-4,6-androstadiene-17fl-ol-3-one is added to each shake flask as a dimethylformamide solution (100 mg./ ml.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The 6 methyl 17oz chloroethynyl 1,4,6 androstatriene-l7 8-ol-3-one, is readily crystallized from the concentrate. Descending paper chromatography of the prodnot in a system using forrnamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A-dehydrogenation product.

Bacillus sphtrericus (ATCC 12488) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Shefiield Farms), 5% glucose, and 0.5% corn steep liquor. After a 24 hour incubation at 28 C., 10 mg. of 6cx-methyl-17u-chloroethynyl-4,6-androstadiene-17l3-ol-3-one is added to each shake flask as a dimethylformamide solution (100 mg./ ml.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The 60: methyl 17cc chloroethynyl 1,4,6 androstatriene-17p-ol-3-one, is readily crystallized from the concentrate. Descending paper chromatography of the product in a system using formamide as the stationary phase, and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

In accordance with the above procedures, but starting with the 6u-methyl l7ix-bromoethynyl-4,d-androstadienel7fi-ol-3-one in place of the 6oi-methyl-17ct-chloroethynyl- 4,6-androstadiene-llfi-ol-one, there is obtained the 6mmethyl 17cc bromoethynyl 1,4,6 androstatriene 17,8- ol-3-one.

Exam ple 9 A mixture of 500 mg. of the 6a-n1ethyl-17wchloroethynyl-4-androstene-17,8-oh3-one, 1'0 of dimethyl formamide, 20 ml. of methyl iodide, and 1.5 gms. of silver oxide is stirred at room temperature for 4 days. An additional /2 gm. of silver oxide is added at the end of each day. One hundred ml. of chloroform is then added to the reaction mixture, then stirred for one hour, filtered,,and the filtrate evaporated to dryness. The residual oil is chromatographed with acid-washed alumina 16 and eluted with mixtures of ether and petroleum ether to give a 6a-methyl-17a-chloroethynyl-4-androstene-17emethoxy-3-one.

In accordance with the above procedures, but starting with the 6u-methy1-17u-bromoethynyl-4-androstene-17pol-3-one in place of the fia-methyl-l7a-chloroethynyl-4- androstene-17fi-ol-3-one there is obtained the 6ct-methyl- 17u-bromoethynyl-4-androstene-17;8-methoxy-3-one.

10!} mg. of 6a methyl-1-7a-chloroethynyl-4-androstene- 17B-ol-3-one is heated with 1. cc. of acetic anhydride and 1.2 cc. of pyridine on the steam bath over night. The reaction mixture is then poured onto ice and extracted with chloroform. The extract is washed with water and concentrated. The concentrate is chromatographed over acid-washed alumina and eluted with mixtures of ether and petroleum ether to give 6a-methyl-17a-chloroethynyl- 4-androstene-l7B-ol-3-one acetate.

In accordance with the above procedures, but starting with the 6m-methyl-17a-bromoethynyl-4-androstene-17B- ol-3-one in place of the 6tz-methyl-17a-chloroethynyl-4- androstene-175-ol-3-one there is obtained the Got-methyl- 17a-bromoethynyl-4-androstene-17l3-0l-3-one acetate.

Example 10 mg. of 17a-chloroethynyl-4-androstene-1718-01-3- one (Example 1) is heated with 1 cc. of acetic anhydride and 1.2 cc. of pyridine on the steam bath overnight. The reaction mixture is then poured onto ice and extracted with chloroform. The extract is washed with water and concentrated. The concentrate is chromatographed over acid-washed alumina and eluted with mixtures of etherpetroleum and ether to give 17a-chloroethynyl-4-androstene-17fi-ol-3-one acetate.

In accordance with the above procedure, but starting with the 17ot-bromoethynyl-4-androstene-17,B-ol-3 one in place of the 17or-chlo'roethynyl-4-androstene-17,9-ol-3-one there is obtained the 17e-bromoethynyl-4-androstene-17dol-3-one acetate.

Example 11 To 100 mg. of 17a-chloroethynyl-4-androstene-17 8-01- 3-one acetate in 5 ml. of t-butanol and 0.1 ml. of acetic acid isadded 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is Washed successively with aqueous sodium bicarbonate, water, dilute hydrochloric acid and Water, and then dried over magnesium sulfate. The extract is treated with activated charcoal and then concentrated to dryness. Crystallization of the residue gives 17a-chloroethyny1-1,4-androstadiene-17fl-ol- 3 one acetate.

In accordance with the above procedures, but starting with the 17m-bromoethynyl-4-androstene-17,8-01 3 one acetate in place of the 17a-chloroethynyl-4-androstene- 17B-ol-3-one acetate, there is obtained the 17a-bromoethynyl-l,4-androstadiene-17fi-ol-3-one acetate.

Corynebacterium simplex (ATCC 6946) is inoculated from a slant to 250 ml. shake flasks containing a medium having the composition: 1 g./liter yeast extract (Difco). After an 18 hour growth phase at 28 C., 10 mg. of 17achloroethynyll-androstene-17B-ol-3-one acetate is added to each flask as a dimethylformamide solution (100 mg./ 1111.). After a 24 hour transformation period at 28 C., the cells are centrifuged, followed by three ethyl acetate extracts of the cell-free broth. The extracts are combined and concentrated in vacuo. The l7u-chloroethynyl- 1,4-androstadiene 17B-ol-3-one acetate is crystallized directly from the concentrate. Paper chromatography of the product in a system utilizing formamide as the stationary phase and chloroform-benzene (1:1) as the mobile phase indicates that the product possesses a polarity slightly greater than the substrate.

Septomyxa ajjims (ATCC 6737) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Shefiield Farms), glucose, 0.5% corn steep liquor. After a 48 hour incubation at 28 C., mg. of 17a-chloroethynyl-4-androstene- 1718-ol-3-one acetate is added to each shake flask as a dimethylformamide solution (100 mg./ml.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The product is re-acetylated by treatment with acetic anhydride and pyridine, and then purified by recrystallization to give the l7a-chloroethynyl-1,4-androstadiene-l75- ol-3-one acetate. Descending paper chromatography of the product in a system using formamide as the stationary phase, and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A-dehydrogenation product.

In accordance with the above procedures, but using the 17owbromoethynyl-4-androstene-175-01- 3 one acetate in place of the 17a-chloroethynyl-4-androstene-17B-ol-3-one acetate, there is obtained the 17a-bromoethynyl-1,4-androstadiene-17fi-ol-3-one acetate.

Example 12 A mixture of 500 mg. of the l7a-chloroethynyl-4-androstene-17/3-ol-3-one, 10 ml. of dimethylformamide, 20 ml. of methyl iodide, and 1.5 gms. of silver oxide is stirred at room temperature for 4 days. An additional A1 gm. of silver oxide is added at the end of each day. One hundred ml. of chloroform is then added to the reaction mixture, which is then stirred for one hour, filtered, and the filtrate is evaporated to dryness. The residual oil is chromatographed with acid-washed alumina and eluted with mixtures of ether and petroleum ether to give 17a-chloroethynyl-4-androstene- 17 ,B-methoxy-3-one.

In accordance with the above procedures, but starting with the 17u-bromoethynyl-4-androstene-17B-ol-3-one in place of the 17a-chloroethynyl-4-androstene-17/3-ol-3one, there is obtained the 17a-bromoethynyl-4-androstene-17B- methoxy-3-one.

Example 13 To 100 mg. of l7a-chloroethynyl-4-andros-tene-17 3 methoxy-3-one in 5 ml. of t-butanol and 0.1 ml. of acetic acid is added 50- mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is washed successively with aqueous sodium bicarbonate, water, dilute hydrochloric acid and water, and then dried over magnesium sulfate. The extract is treated with activated charcoal and then concentrated to dryness. Crystallization of the residue from a mixture of acetone and ether gives 17oc-Cl1l010- ethynyl-1,4-androstadiene-l7/3-methoxy-3-one.

Septomyxa afiinis (ATCC 6737) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Sheflield Farms), 5% glucose, and 0.5% corn steep liquor. After a 48 hour incubation at 28 C., 10 mg. of l7a-chloroethynyl-4-andr0stene-17fi-methoxy-3-one is added to each shake flask as a dimethylformamide solution (100 mg./ml.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The 17u-chloroethynyl-1,4-androstadiene-17/3-methoxy- 3-one is readily crystallized from the concentrate. Descending paper chromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

Bacillus sphaericus (ATCC 12488) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Shefiieid Farms), 5% glucose, and 0.5% corn steep liquor. After a 24 hour incubation at 28 C., 10 mg. of 17achloroeth-ynyl- 4 androstene-l7fl-methoxy-3-one is added to each shake flask as a dimethylformamide solution mg./rnl.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The l7ot-chloroethynyl-l,4-androstadiene-l7fi-methoxy- 3-one, is readily crystallized from the concentrate. De scending paper chromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is langely the somewhat more polar A dehydrogenation product.

In accordance with the above procedures, but using the 17a-bromoethynyl-4-androstene-17,8-methoxy 3 one in place of the 17a-chloroethynyl-4-androstene-17/8-methoxy- 3-one, there is obtained the 17a-hromoethynyl-l,4-androstadiene-l7fi-methoxy-3-one.

Example 14 A mixture of 1 g. of l7or-chloroethynyl-4-androstenel7,B-ol-3-one, 10 ml. of acetic anhydride and 100 mg. of p-toluenesulfonic acid is heated on the steam bath for one hour and allowed to stand overnight at room temperature. It is then poured into ice Water and left to stand at room temperature for one half hour. The reaction mixture is then extracted with ether. The ether extract is washed with water and aqueous sodium bicarbonate, dried, and concentrated to yield the crude 17achloroethylnyl-3,5-androstadiene-3,17,8-diol diacetate. The product, crystallized from petroleum ether, has a melting point of -131 C. The analysis of a sample which has a melting point of -l36 C. is: C, 69.75; H, 7.23. Calculated C, 69.67; H, 7.25.

To a solution of 200 mg. of the l7u-chloroethyn-yl-3,5- androstadiene-S,l7)8-:diol-diacetate dissolved in 2 ml. of acetic acid is aclded with stirring, 56 ml. of N-chlorosuccinimide and 2 ml. of tetrahydrofuran containing 5% of dry HCl. The reaction is stirred at room temperature for 2 /2 hours. Sodium bicarbonate is added and the reaction mixture is extracted with ether. The ether extracts are dried and concentrated. The material is chromatographed over acid-Washed alumina, and eluted 'With ether-petroleum ether mixtures, to give the 6u-chloro- 17a-chloroethynyl-4-androstene-l7}8-ol-3-one acetate. The analytical sample is crystallized from a mixture of methylene chloride-ether and has the following properties: M.P. 220-230 C.

an... 235 m,., 6 13,600

Analysis.Calculated: C, 65.26; H, 6.67; Cl, 16.75. Found: C, 65.80; H, 6.74; Cl, 17.86.

In accordance with the above procedures, but starting with the 17abromoethynyl-4-androstene-17/3-ol-3-one in place of the 17wchloroethynyl-4-androstene-17B-ol-3-0ne, there is obtained 'as intermediate, the l7a-b-romoethynyl- 3,5-androstadiene-3,l7B diol-diacetate, and as product, the 6a-chloro-17owbromoethynyl-4-androstene-1718-01- 3 one acetate.

Example 15 To 100 mg. of 6a.chloro-17a chloroethynyl-4-androstene-l7B-ol-one acetate in 5 ml. of t-butanol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is washed successively with aqueous sodium bicarbonate, ammonium sulfide, dilute ammonia water, water, dilute hydrochloric 19 acid and water, and then driedv over magnesium sulfate. The extract is treated withz:.activated charcoal and then concentrated to. dryness. Crystallization of the residue from a mixture of acetone and ether gives 60z-Ch1O1'0-17achloroethynyl-1,4-androstadiene-17,8-ol-3-one acetate.

In accordance with the above procedures, but starting with the 6a-chloro-l7u-brornoethynyl-4-androstene-17 3-01- 3-one acetate in place of the Got-chloro-17m-chloroethynyl- 4-androstene-17fl-ol-3-one acetate there is obtained the 6a-chloro-17a-bromoethynyl-1,4-androstadiene-l7fi-ol 3- one acetate.

. Corynebacteriltm simplex (ATCC 6946) is inoculated from a slant to 250 ml. shake flasks containing a medium haying .the composition: .1 .gJliter yeast extract (Difco). After an 18 hour growthphase at 28 C., 10 mg. of

6a-chloro-l7wchloroethynyl 4-androstene-l7B-ol-3-one slant into 250ml. shake flasks containing 50 ml. of the following medium: 2% Eda-min (Shefiield Farms), 5% glucose, and 0.5% corn steep liquor. After a 48 hour incubationat 28 C., mg. of 6a-chloro-17achloroethynyl-4-androstene-17 3-ol-3-one acetate is added to each shake flask-as a dimethylformamide solution (100 mg/ ml.). After a conversion period of 24-hours, the cells are removed by filtration, followed by three successive extractionswith equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The product is re-acetylated by treatment with acetic anhydride and pyridine, and then purified by recrystallization to give the 6a-chloro-lhchloroethynyl-lA-andro-- stadiene-17y8-ol-3-one acetate, is readily crystallized from the concentrate.

phase and chloroform as the mobile phase, shows some of the steroid substrate, but is langely the somewhat more polar Adehydrogenation product.

Descending paper chromatography of the product in a system using formamide as the stationary In accordance with the above procedures,but using the 6a chloro 17a bromoethynyl 4 androstene- 17,3- ol-3-one acetate in place of the Got-ChlOl'O-l'loc-ChlOIO- ethynyl-4-androstene-17fl-ol-3-one acetate, there is obtained the 6u-chloro-17a-bromoethynyl-1,4-iandrostadi Y ene-l7 8-ol-3-one acetate.

Example 16 Amixture'oi 1 got 17c-chloroethynyl-4-androstene-.

17fl-methoxy-3 one (Example .10), 10ml. of acetic anhydride and 100 mg. of p-toluenesulfonic acidis heated on the steam bath for one hour and allowed to. stand overnight at room temperature. It is then poured into ice water and left tostand at room temperature forone half hour. The reaction mixture isthen extracted with ether.

The ether extract is washed with water and aqueous. so-

dium bicarbonate, dried, and concentrated to yield the 17d chloroethynyl 3,5 androstadiene 17,6 methoxy- 3-ol acetate.

To a solution of 200 mg. of the 17e-chloroethynyl-3,5- androstadiene-17fl-rnethoxy-3-'olacetate dissolvedin 2 ml. of acetic acid is added, with stirring, 56 ml. of N-chlorosuccinimide and 2 ml. of tetr-ahydrofuman containing 5% of dry HCl. The reaction is stirred at room temperature for 2 /2 hours. Sodium bicarbonate is added... The reaction mixture is then extracted with ether.. The ether extracts are dried and concentrated. The material.-.is chromatographed. over acid-washed alumina, an-d=eluted 2'0 with ether-petroleum ethermixtures to give Get-chlorol7u-chloroethynyl-4-androstene 17fi-methoxy-3-one.

In accordance with the above procedures, but starting with the l7a-bromoethynylA-andr'ostene-17B-methoxy-3- one in place of the l7u-chloroethynyl-4-androstene-l7,6- methoxy-3-one, there is obtained as intermediates, the 17m chloroethynyl 3,5 androstadiene 17,8 methoxy- 3-ol-acetate, and as product, the 6at-chloro-l7u-chloroethynyl-4-androstene-l7B-methoxy-3-one.

Example 17 Tov mg. of 17-a-chloroethynyl-4-androstene-17B- methoxy-B-one in 5 mlrof t-but-anol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue isextracted with ethyl acetate and the extract is washed successively with aqueous sodium bicarbonate, ammonium sulfide, diluteammonia water, water, dilute hydrochloric acid and water, and then dried over magnesium sulfate. The extract is treated with activatedcharcoaland then concentrated to dryness. Crystallization of the residue from a mixture of acetone and ether gives \17a-chloroet-hynyl-l, 4-androstadienel 7fl-methoxy-3-one.

Septomyxa afiinis (ATCC 6737) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Sheflield Farms), 5% glucose, and 0.5% corn steep liquor. After a 48 hour incubation at 28 C., 10 mg. of l7a-chloroethynyl-4- androstene--methoxy-3-one is added to each shake flask as a dimethylformamide solution (100 mg./-ml.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. are combined and concentrated in vacuo.

The l7dtchloroethynyl-1,4-androstadiene-l713-methoxy- 3-0ne is'readily crystallized from the concentrate. Descending paper chromatography of the product in a system using formamide as the stationary phase and chloro-' form as the mobile phase, shows some of the steroid substrate, but 'is' largely the somewhat more polar A-dehydrogenation product.

Bacillus .s'phaericus (ATCC 12488) is inoculated from a slant into 250ml. shake flasks containing 50 ml. of thefollowing medium: 2% Edamin (Sheffield Farms), 5% glucose, and 0.5% corn steep liquor. Aftera 24 hour incubation at 28 CI, 10 mg. of l7a-chloroethynyl-4- androstene-l7 8-methoxy-3-one is added to each shake flask as a dimethylformamide solution (100 mg.'/ml.). After a conversion period of 24 hoursythe cells are" aremoved by-filtration, followed-by three successive extractions with equal volume of ethylacetate. The extracts are combined and concentrated in vacuo.

The l7u-chloroethynyl-1,4-androstadiene-17B-methoxy- 3-one islreadily crystallized from the concentrate. Descending paper chromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

In accordance with the above procedures, but using'the 17a-bromoethynyi-4-androstene 175 methoxy-3-one in place of the l7a-chloroethynyl-4-androstene l7B-methoxy-3-one, there is obtained-the 17a-bromoethynyl-1,4-'

androstadiene- 17 flmethoxy-3-one.

Example 18 A suspension of 1 g. of 17u-chloroethynyl-4-androstene- 17 8-ol-3-one and 2.4 g. of chloranil in 30 ml. of ethyl acetate and 6 ml. of acetic acid is stir-red and refluxed for 16 hours. The reaction mixture is cooled and filtered. The product is washed sequentially with aqueous bisulfite, aqueous pot-assium'hydroxide and water.

The extracts Chroma 21 tography yields 17a-chloroethynyl-4,6-andr0stadiene-17B- ol-3-one which on crystallization from ethyl acetate has the following properties: M.P. 2092ll C.: 11 -l59 (c. 0.9 CHCl U.V. A3??? 283 mu, 6 26,900

Calculated for C H O Cl; C, 73.15; H, 7.31. Found: C, 73.08; H, 7.06.

A solution of 675 mg. of 17a-chloroethynyl-4,6-androstadiene-l7fl ol-3-one in 30 ml. of ether and 30 ml. of 0.30 N perbenzoic acid in benzene is allowed to stand in the dark for 68 hours. The solution is washed sequentially with aqueous sodium bisulfite, water, 2.5 N potassium hydroxide and water. The organic layer is dried and concentrated in vacuo. Chromatography on acid-washed alumina and elution with ether aflords about 180 mg. of the 17ot-chloroethynyl-4-androstene-l7fi-ol-3-one-6,7-oxide, which has the following properties: M.P. 196216 C.

U.V. A1552 240 m 13% 377 A solution of 140 mg. of the oxide in 10 ml. of 0.4 N hydrogen chloride in chloroform is allowed to stand at room temperature for 5.5 hours. The solution is poured onto ice. Aqueous sodium bicarbonate solution is added and the product is extracted with chloroform. The organic layer is washed with water, dried and concentrated. Crystallization from ether afiords 84 mg. of 6-chloro-17 chloroethynyl-4,6-androstadiene-17,3-ol-3-one, M.P. 19 205 C. The sample for analysis is crystallized from methanol and has the following properties: MP. 203-- 208" 0.; 0: 96 (c. 1.0 CHCl U.V. A355? 285 my, 6 21,400

Calculated for C H O Cl C, 66.49; H, 6.38. Found: C, 66.28; H, 6.62.

In accordance with the above procedures, but starting With the l7a-bromoethynyl-4-androstene l7,B-olr3-one in place of the l7a-chloroethynyl-4-androstene-17j8-ol-3-one, there is obtained as intermediate, the l7ot-bromoethynyl- 4-androstene-17fl-ol-3-one-6,7-oxide, and as product, the 6-chloro-17-bromoethynyl-4,6-androstadiene-l7B-ol-3-one.

A solution of 125 mg. of 6-chloro-l7a-chloroethynyl- 4,6-androstadiene-17B-ol-3-one in 1.8 ml. of pyridine and 1.5 ml. of acetic anhydride is heated on the steam bath overnight. The solution is poured into ice water and extracted with ether. The ether solution is Washed sequentially with dilute hydrochloric acid, water and sodium bicarbonate solution. Chromatography on 7.0 g. of acidwashed alumina, and elution with ether; petroleum ether mixtures affords 56 mg. of 6-chloro-l7a-chloroethynyl- 4,6-androstadiene-17,8-01-3-one acetate. The sample tor analysis is crystallized from ether and has the following properties: M.P. 203-208 C., 0: 98 (c. 0.9 CHCl U.V. urg 284 6 21,300

Calculated for C H O Cl C, 65.57; H, 6.21. Found: C, 66.06; H, 6.16.

In accordance with the above procedures, but starting with the 6 dhloro-17a-bromoethynyl-4,6-androstadiene- 17;3-ol-3-one in place of the 6-chloro-17u-chloroethynyl- 4,6-androstadiene-l7fl-ol-3-one, there is obtained the 6- chloro 17a-bromoethynyl-4,6-androstadiene-17B-ol-3-one acetate.

Example 19 To 100 mg. of 6u-chloro-17a-chloroethynyl-4,6-androstadiene-175-ol-3-one acetate in 5 ml. of t-butanol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is washed successively with aqueous sodium bicarbonate, water, dilute hydrochloric acid and water, and then dried over magnesium sulfate. The extract is treated with activated 22 charcoal and then concentrated to dryness. Crystallization of the residue gives 6-chloro-17a-chlor0ethynyl-l,4,6- androstatriene-17/8-ol-3-one acetate.

Corynebacterium simplex (ATCC 6946) is inoculated from a slant to 250 ml. shake flasks containing a medium having the composition: 1 g./liter yeast extract (Difco). After an 18 hour growth phase at 28 C., 10 mg. of 6- chloro 17a-chloroethynyl-4,6-androstadiene-175-ol-3-one acetate is added to each flask as a dimethy-lformamide solution rug/ml). After a 24 hour trans-formation period at 28 C., the cells are centrifuged, followed by three ethyl acetate extracts of the cell-free broth. The extracts are combined and concentrated in vacuo. The 6- ohloro l7a-chloroethynyl-1,4,6-androstatriene-17501-3- one acetate is crystallized directly the concentrate. Paper chromatography of the product is a system utilizing tormamide as the stationary phase and chloroform-benzene (1 :1) as the mobile phase indicates that the product possesses a polarity slightly greater than the substrate.

Septomyxa aflinis (ATCC 6737) is inoculated from a slant into 250 ml. shake flasks containing 50 m1. of the following medium: 2% Edamin (Sheffield Farms), 5% glucose, and 0.5% corn steep liquor. After a 48 hour incubation at 28 C., 10 mg. of -6-chloro-17u-chloroethynyl-4,6-androstadiene-17/8-ol-3-one acetate is added to each shake flask as a dimethylformamide solution (100 mg./ml.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The product is re-acetylated by treatment with acetic anhydride and pyridine, and then purified by recrystallization to give the 6-chloro-17a-chloroethynyl-1,4,6-androstatriene-l7fi-ol-3 one acetate. Descending paper chromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

In accordance with the above procedures, but using the 6 chloro-17a-bromoethynyl-4,6-androstadiene-175 01-3- Example 20 A mixture of 500 mg. of 6-chloro-17u-chloroethynyl- 4,6-androstadiene-17B-ol-3-one, l0 of dimethylformamide, 20 ml. of methyl iodide, and 1.5 gms. of silver :oxide is stirred at room temperature ior 4 days. An additional /2 gm. of silver oxide is added at the end of each day. 100 ml. of chloroform is then added to the reaction mixture Which is then stirred for one hour, filtered. The filtrate is then taken to dryness. The oil is chromatographed with acid-washed alumina and eluted with mixtures of ether and petroleum ether to give the 6-chloro- 17u-chloroethynyl-17/8-meflhoxy-4,6-androstadiene-3-one.

In accordance with the above procedures, but starting with the 6-chloro-17a-bromoethynyl-4,G-androstadiene- 17B-ol-3-one in place of the 6-chloro-l7a-chloroethynyl- 4,6-androstadiene-17B-ol-3-one there is obtained the 6- chloro-17a-bronroethynyl-17fi-methoxy-4,6 androstadiene- 3-one.

Example 21 To 100 mg. of 6-chlor0-l7a-chloroethynyl-17,8-methoxy-4,6-androstadiene-3-one in 5 ml. of t-butanol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is Washed successively with aqueous sodium bicarbonate, ammonium sulfide, dilute ammonia Water, Water, dilute hydrochloric acid and water, and then dried over magnesium sulfate.

23 The extract is treated with activatedcharcoaland then concentrated to dryness. Crystallization of the residue from a mixture of acetone and ether gives the 6-chloro- 17a-chloroethynyl- 1713 metho'xy-1,4,6 androstatriene-3- one.

Sepzomyxa ajfinis (ATCC 6737) is inoculated frorna slant into 250 ml. shake flasks containing 50 ml.'- of the" following medium: 2% Edamin (Sheifield Farms), 5% glucose, and 0.5% corn steep liquor; After a 48-hour incubation at 28 C., 10 mg. of 6-ch1oro-17a-chloroethy'nyl-17,8-methoxy-4,6-androstadiene-3-one is added to eachshake'flask as a dimethylformamide solution" (100 sing/ml). After a conversion'period of 24'hours, the cells areremoved by filtration, followed by three" successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The '6-chloro-17a chloroethynyl-17B methoxy 1,4,6- andro'statriene-3-on'e is readily crystallized'from the concentrate'. Descending paper chromatography of-the produc-tin a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat'more polar" A'-dehydrogenation product.

Bacillus sphaericus (ATCC 12488) is inoculated froma slant into 250 ml. shake flaskscontaining 50 ml. ofthe following-medium: 2% Edamin (Sheffield-Farms), 5% glucose, and'*0.5% cornsteep'liquor. After a 24 hour incubation'at 28 (3., 10 mg. of6 chloro-l7m-chloroethyny1-4,6"-androstadiene-17B-methoxy-3-orie" is added to' each shake flask" as a dimethylfornramide solution (100 mg./ml.'). After a conversion period of 24'hours',=the cells are removed byfiltration, follow'e'd by three successive'extractioris with equal volume' of'ethylacetate: The extracts are combined and concentrated inva'c'uo;

The 6-chloro-l7ot-chloroethynyl 1713 mthoxy 1,4,6- andorstatriene-3-one "is readily crystallized from-the 'con-' centrate. Descendingpaper chromatography of-the prod not in a system usingfor-mamide as the-stationary phaseand chloroform asthe mobile *phase,--'shows someof the steroid substrate, but is largely the somewhat more polar- A'-dehydrogenation product.

. In accordance with the above procedures,-but-using the" 6-chloro-17a-bromoethynyl-4,6-androstadiene 17B'- methoxy-3-one in place of the 6-chloro-17u-chloroethynyl-4,6- androstadiene-l7fi-methoxy-3-one; there is obtained the 6-chloro-17e brdmoethynyl 1,4,6 andi'o'statriene 17 18- methoxy-3-one.

ExampIeZZ Asolution of the 17ot-chloroethynyl-fi,S-and'roStadiene- 313,175-diol diacetate (Example 10) in aqueouste'trahydrofu'ran is treated with a slow stream oi perchloryl fluoride for one hourand then allowed to stand at room temperature for an additional 2 hours. The reaction inixture is poured on ice and extracted with chloroforrnfl The chloroform layer is washed with aqueous bicarbonate solution, dried and concentrated in vacuo. The crude material is dissolved in acetic acid saturated with gaseous hydrogen chloride and allowed to s-tandifor "one hour at room temperature. The reaction mixture is poured on ice and extracted with'chloroform. The chloroform layer' is Washed with aqueous bicarbonate' solution, dried and concentrated in vacuo. The concentrate is chroma-t graphed on acid-washed alumina-to yield the 6a-fluoro- 17a-chloroethynyl-4-androstene-17 8-01-3-one acetate.

In accordance with the above procedures, but starting with the 17a-bromoethynyl-3,S-androstadiene-3B,17fl-diol diacetate in place of the 17a-chloroethynyl-3,5-androstadicue-3,8,17,8-diol diacetate there is obtained the tied-fluoro- 17a-bromoethynyl-4-androstene-l7fi-ol-3-one acetate.

Example 23 To 100 mg. of 6u-fluoro-17a-chloroethynyl-4-androstene-17fi-ol-3-one acetate in ml. of t-butanol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 24 50 rng. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is Washed suc- 5' cessively with aqueous sodium bicarbonate, water, dilute hydrochloric acid and water, and then dried over magnesium sulfate. The extract is treated with activated charcoal and then concentrated to dryness. Crystallization of the residue from a mixture of acetone and ether gives 6u-fluoro-17a-chloroethynyl-1,4-androstadiene-17B- ol-3-one actate.

I Corynebacterium simplex (ATCC 6946) is inoculated from a slant to 250 ml. shake flasks containing a medium having the composi-tioni 1 g./liter yeast extract (Difco).

1 After an 18-hour growth phase at 28 C., 10 mg. of 6afiuoro-l7:x-chloroethynyl-4-androstene-17,8-ol-3-one acetate is added to each flask as a dirncthylformamide solution (100 mg./ml.). After a 24-hour transformation period at 28 C., the cells arecentrifuged, followed by three ethyl acetate extracts of the cell-free broth. The extracts are combined and concentrated in vacuo- The 6-fluoro-17a-chloroethynyl-1,4- androstadiene 175-01-3- onea'ctate is crystallized directly fromthe concentrate. Paper 'chrdmatography of the product is a system utilizing formamide as the stationary and chloroform-benzene (1:1) as the mobile phase indicates that the product possesses a polarity slightly greater than the substrate.

Septdmyxa'afiihis (ATCC 6737) is inoculated from a slant iinto'250 ml. shake flasks containing 50 mliof the 30 followingmedium: 2% Edamin(Shefiield Farms), 5% glucose, and 0.5% cornsteep liquor. After a '48 hour incubation at 28 C., 10 mg. of 6a-fluoro-17u-chloroethynyl-{l-androstene-1713-01-3-one acetate is added to each shake flask as a dimethylformamide solution (100 mg./

ml.) After a conversion period of 24 'hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The product is re-acetylated by treatment with acetic anhydride and pyridine, and then purified by recrystallization to give the a-fluoro-l7a-chloroethynyl-1,4-androstadiene-l7fi-ol-3-one acetate. Descending paperchromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, "shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

In accordance with the above procedures, but using the fiuoro 17ot-bromoethynyl-4-androstene-l7,8-0l-3-one acetate in place of the fia-fiuoro-l7a-chloroethynyl-4-an- "drostene47B-ol-3-one acetate there is obtained the 6mfluo ro l7a-bromoe thynyl-1,4-androstadiene-17fl-ol-3-one acetate.-

Example 24 Example 25 Tci 100 mg. of 6-fluoro-17a-chloroethynyl-4,6-androstadiene-17B-ol-3-one acetate in 5 ml. of 't-bu tanol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide.

. The mixture is refluxed under nitrogen for 18 hours; then 50' mg; of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is washed successively with aqueous sodium bicarbonate, water, dilute hydrochloric acid and water, and then dried over magnesium sulfate. The extract is treated with activated charcoal and then concentrated to dryness. Crystallization of the residue gives 6-fluoro-17a-chloroethynyl-1,4,6-androstaniene-17 3-ol-3-one acetate.

In accordance with the above procedures, but starting with the 6-fluoro-17u-bromoethynyh4,6-androstadiene-l7fi-ol-3-one acetate in place of the 6-fluoro-17achloroethynyl 4,6 androstadiene-l7B-ol-3-one acetate, there is obtained the 6-fluoro-17ot-bromoethynyl-1,4,6- androstatriene-l7fi-ol-3-one acetate.

Corynebacterium simplex (ATCC 6946) is inoculated from a slant to 250 ml. shake flasks containing a medium having the composition: 1 g./liter yeast extract (Difco). After an 18 hour growth phase at 28 C., 10 mg. of 6 fluoro-17a-chloroethynyl-4,6-androstadiene-175-01-3- one acetate is added to each flask as a dimethylformamide solution (100 mg./ml.). After a 24 hour transformation period at 28 C., the cells are centrifuged, followed by three ethyl acetate extracts of the cell-free broth. The extracts are combined and concentrated in vacuo. The 6-fluoro-17a-chlloroethynyl1,4,6-androsta|triene-17B- ol-3-one acetate is crystallized directly from the concentrate. Paper chromatography of the product in a system utilizing formamide as the stationary phase and chloroform-benzene 1:1) as the mobile phase indicates that the product possesses a polarity slightly greater than the substrate.

Septomyxa afiznis (ATCC 6737) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Sheffield Farms), 5% glucose, and 0.5% corn steep liquor. After a 48 hour incubation at 28 C., mg. of 6-fluoro-17a-chloroethynyl-4,6-androstadiene-17fl ol-3-one acetate is added to each shake flask as a dimethylformamide solution (100 mg./ml.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The product is re-acetylated by treatment with acetic anhydride and pyridine, and then purified by recrystallization to give the 6ot-fluoro-l7oc-chloroethynyl-1,4,6- androstatriene-17fi-ol-3-one acetate. Descending paper chromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but it is largely the somewhat polar A-dehydrogenation product.

In accordance with the above procedures, but using the 6-fluoro-l7a-bromoethynyil-4,6- androstadicne-17 3-01- 3-one acetate in place of the 6-fluoro-17a-chloroethynyl- 4,6-androstadiene-175-01-3-one acetate, there is obtained the 6-fluoro-l7oa-bromoethynyl-1,4,6-androstatriene-l7flol-3-one acetate.

Example 26 A solution of the 17a-chloroethynyl-3,S-androstadiene- 17B-methoxy-3fl-ol in aqueous tetrahydrofunan is treated with a slow stream of perchloryl fluoride for one hour and then allowed to stand at room temperature for an additional 2 hours. The reaction mixture is poured on ice and extracted with chloroform. The chloroform layer is washed with aqueous bicarbonate solution, dried and concentrated in vacuo. The crude material is dis solved in acetic acid saturated with gaseous hydrogen chloride and allowed to stand for one hour at room temperature. The reaction mixture is poured on ice and extracted wtih chloroform. The chloroform layer is washed with aqueous bicarbonate solution, dried and concentrated in vacuo. The concentrate is chromato- 26 graphed on acid-washed alumina to yield the GOL-flUOI'O- 17a-ch1oroethynyl-4-androstene-17,8-methoxy-3-one.

In accordance with the above procedures, but starting with the 17a-bromoethynyl-3,S-androstadiene-17,8-methoxy-3p-ol in place of the 17a-chloroethynyl3,S-androstadiene-17fl-methoxy-3/3-ol, there is obtained the 6a-fluoro- 17a-bromoethynyl-4-androstene-17fl-methoXy-3-one.

Example 27 To mg. of 6a-fluoro-17a chl-oroethynyl-4-androstene-17fl-methoxy-3-one in 5 ml. of t-butanol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is eX- tracted with ethyl acetate and the extract is Washed successively with aqueous sodium bicarbonate, water, dilute hydrochloric acid and water, and then dried over magnesium sulfate. The extract is treated with activated charcoal and then concentrated to dryness. Crystallization of the residue gives 6a-fluoro-l7a-chloro-ethynyl-1,4-androstadiene-l7B-methoxy-3-one.

Septomyxa afinis (ATCC 6737) is inoculated from a slant into 250 ml. shake flasks containing 50 of the following medium: 2% Edamin (Sheflield Farms), 5% glucose, and 0.5% corn steep liquor. After a 48 hours incubation at 28 C., 10 mg. of 6a-fluoro-17a-chloroethynyl-4-androstene-l7B-methoxy-3-one is added to each shake flask as a dimethylfonnamide solution (100 mg./ ml.). After a conversion period of 24 hours, the cells are removed :by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The 6u-fluoro-17ot-chloroethynyl-1,4-androstadiene-17/3- methoxy-3-one is readily crystallized from the concentrate. Descending paper chromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

Bacillus sphaericus (ATCC 12488) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Shefiield Farms), 5% glucose, and 0.5% corn steep liquor. After a 24 hour incubation at 28 C., 10 mg. of 6oc-fi-UOTO-17cc-Chl0l'0- ethynyl-4-androstene-17fi-methoxy-3-one is added to each shake flask as a dimethyl-formamide solution (100 mg./ ml.). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The 6a-fluoro-17a-chloroethynyl-l,4-androstadiene-l7,8- methoxy-3-one is readily crystallized from the concentrate. Descending paper chromatography of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

In accordance with the above procedures, but using the 60c fluoro 17oz ibromoethynyl 4 androstene 17,8- methoxy-3-one in place of the 6a-fluoro-17a-chloroethyny1 4-androstene-17fi-methoxy-3-one, there is obtained the 6:1- fluoro .170 bromoethynyl 1,4 androstadiene 17B- methoxy-3-one.

Example 28 A suspension of 1 g. of Got-*fillOIO-17oc-Cl1l01'06thYI1Yl- 4-androstene-l7,B-rnethoxy-3-one, 2.4 g. chloranil, 30 ml. ethyl acetate and 6 m1. of acetic acid is stirred and refluxed for 16 hours. The reaction mixture is cooled and filtered. The product is washed sequentially with aqueous hisulfite, aqueous potassium hydroxide and water. The organic layer is dried andconcentrated in vacuo. Chro 27 matography yields 6-fiuoro-17a-chloroethynyl-4,-androstadiene-l7B-rnethoxy-3-one.

Example 29 To 100 mg. of 6-fluoro-17oc-chloroethynyl-4,6-androstadiene-l7fi-methoxy-3-one in ml. of t-butanol and 0.1 ml. of acetic acid is added 50 mg. of selenium dioxide. The mixture is refluxed under nitrogen for 18 hours; then 50 mg. of selenium dioxide is added and the mixture is refluxed an additional 24 hours. The product is filtered, and the filtrate evaporated to dryness. The residue is extracted with ethyl acetate and the extract is washed successively with aqueous sodium bicarbonate, water, dilute hydrochloric acid and water, and then dried over magnesium sulfate. The extract is treated with acetivated charcoal and then concentrated to dryness. Crystallization of the residue gives 6-fluoro-l7a-chloroethynyl- 1,4,6-androstatriene-l7fi-methoxy-3-one.

Septomyxa alfinis (ATCC 6737) is inoculated from a slant into 250 ml. shake flasks containing 50 ml. of the following medium: 2% Edamin (Shefiield Farms), 5% glucose, and 0.5% corn steep liquor. After a 48 hour incubation at 28 C, mg. of 6fiuoro-17e-chloro ethynyl-4,fi-androstadiene-l7B-methoxy-3-one is added to each shake flask as a dimethyl-for-mamide solution (100 rug/ml). After a conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The 6 fluoro 17w chloroethynyl 1,4,6 androstatriene-17fi-methoxy-3-one is readily crystallized from. the concentrate. Descending paper chromatography of the product in a system using tormamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

Bacillus sphaericus (ATCC 12488) is inoculated from a slant into 250 ml. shake flasks containing 50 of the following medium: 2% Edamin (Sheflield Farms), 5% glucose, and 0.5% cor-n steep liquor. After a 24 hour incubation at 28 0., 10 mg. of 6-fluoro-17e-chloroethynyl-4,6-androstadiene-l7fi-methoxy-3-one is added to each shake flask as a dimethylformamide solution (100 mg./ml.). Afte ra. conversion period of 24 hours, the cells are removed by filtration, followed by three successive extractions with equal volume of ethyl acetate. The extracts are combined and concentrated in vacuo.

The 6 fluoro 17cc chloroethynyl 1,4,6 androstatriene-l7,8-methoxy-3 one is readily crystallized [from the concentrate. Descending paper chromatography of of the product in a system using formamide as the stationary phase and chloroform as the mobile phase, shows some of the steroid substrate, but is largely the somewhat more polar A'-dehydrogenation product.

In accordance with the above procedure, but using the 60: fluoro 17oz bromoethynyl 4,6 androstadiene- 17B-rnethoxy-3-one in place of .the 6a-fluoro-17a-chloroethynyl-4,6-androstadiene-17fi-methoxy-3-one, there is obtained the 6 fluoro- 17cc -bromoethynyl-l,4,6-androstatriene-l7fi-methoxy-3-one.

Example 30 A solution of 100 mg. of the 17m-ehloroethynyl-4-an- 'drostene-l7B-ol-3bne and 50 mg. of Lindlar catalyst in 10 cc. of ethyl acetate is treated with hydrogen until one mole of hydrogen has been absorbed. The mixture is filtered and concentrated. Chromatography yields the pure product 21-chloro-4,20-pregnadiene-175-01-3-one.

In accordance with the above procedures, but starting with the 17a-bromoethynyl-4-androstene-17 8-ol-3-one in place of the 17a-chloroethynyl-4-androstene-17B-ol-3-one there is obtained the 21-chloro-4,20-pregnadiene-l7fi-ol-3- one.

A suspension of platinum oxide in 10 cc. of ethanol is reduced and 100 mg. of 17ot-chloroethynyl-4-androstene- 17 3-ol-3-one is added. Reduction proceeds until two moles of hydrogen have been absorbed. The solution is filtered, concentrated and chromatographed on alumina to yield the 21=chloro-4-pregnen-17,6-o13-one.

In accordance width the above procedures, but starting with the l7ot-bromocthynyl-4-androstene-17,8=ol-3-one in place of the 17a-chloroethynyl-4-androstene-17 8-ol-3-one there is obtained the 2l-ch-loro-4-pregnene-l7fi-ol-3-one.

Example 31 A mixture of 1 g. of 17oc-chloroethynyl-19-nor-4-andro stene-l7/8-ol-3-one, 10 ml. of acetic arrhydride and 100 mg. of p-toluenesu-lfonic acid is heated on the steam bath for one hour and allowed to stand overnight at room temperature. It is then poured into ice water and left to stand at room temperature :for one half hour. The reaction mixture is then extracted with ether. The ether extract is washed with water and aqueous sodium bicarbonate, dried, and concentrated to yield the crude 17a-chloroethynyl-19-nor-3,5-androstadiene-3,l7g3-diol diacetate.

A solution of 8 gm. of 17u-chloroethynyl-l9-nor-3,5- androstadiene-3,l7/8-diol diacetate in a mixture of 700 ml. of ethanol and 300 ml. of tetrahydrofuran is cooled to 10 C. and added dropwise, with occasional stirring, over a one hour period, to a cold (0 C.) solution of 18 g. sodium borohydride in 400 ml. of 80% ethanol, the reaction temperature not being allowed to exceed 5 C. After completion of the addition, the solution is held at 0-5 C. for an additional two hours. A solution of NaH PO is then added to adjust the pH to about 5. The mixture is then concentrated in vacuo to a small volume, diluted with water and extracted with chloroform. The extract is washed with water, dried and concentrated. The concentrate is chromatographed over acid-washed alumina and eluted with ether-petroleum ether mixtures to give 17a-chloroethynyl-19-nor-S-androstene-3fi,l7fl-diol 17-acetate.

Ten grams of l7a-chloroethynyl-l9-nor-5-androstene- 3,8,l7fl-diol-l7-acetate in 150 ml. of tetrahydrofuran is treated with ml. of 0.9 m. monoperphthalic acid in ethyl acetate and allowed to stand over night at room temperature. The reaction mixture is diluted with ethyl acetate and washed sequentially with aqueous sodium bicarbonate, sodium bisulfite, and sodium bicarbonate solution. The organic layer is dried and concentrated in vacuo. The product is fractionally crystallized from aqueous ethanol to yield the 17a-chloroethynyl-l9-norandrostane-3p,17fl-diol-5,6a-oxide-17-acetate.

Five grams of 17a-chloroethynyl19-nor-androstane-3fi, 17fi-diol-5,6a-oxide-l7-acetate in benzene is added to a reagent prepared from 3.4 g. of magnesium, 10 ml. of methyl iodide and 45 ml. of ether. The mixture is stirred and 70 ml. of solvent is removed by distillation. After refluxing for 5 hours, the mixture is cooled, acidified with dilute hydrochloric acid and the organic layer washed to neutrality. The organic phase is dried and concentrated in vacuo. Crystallization from aqueous ethanol gives the 6/3-methyl 17 chlorethynyl-l9-nor-androstane-3fi,5a, 17/i-triol.

The oxidizing reagent is prepared by diluting 2.7 g. of C 0 and 2.3 ml. of concentrated sulfuric acid to 10 ml. with water. A solution of 1.90 g. of GB-methyl-lhchloroethynyl-l9-nor-androstane-6l9,5m,171S-triol in 300 ml. of acetone is cooled to 0 and treated with 1.85 ml. of the oxidizing reagent. The solution is diluted with ice water and extracted with ether. The ether extract is washed sequentially with water, and aqueous sodium bicarbonate and then dried and concentrated in vacuo, to give 6,6-rnethyl-l7a-chloroethynyl-l9-nor-androstane-5a, l7 3-diol-3-one. The crude product, dissolved in ml. of methanol, 75 ml. of 1 N sodium hydroxide is added and the mixture is stirred at room temperature under nitrogen for 24 hours. The mixture is concentrated to half volume under vacuo at 25 C. and then poured into ice water. The product is extracted with ether. The ether layer is washed with water, dried and concentrated. 'Ihe Example 32 100 mg. of 6a-methyl-Hot-chloroethynyl-l9-nor-4-androstene-l7 8-o-l-3-one is heated with 1 cc. of acetic anhydride and 1.2 cc. of pyridine on the steam bath overnight. The reaction mixture is then poured onto ice and extracted with chloroform. The extract is washed with water and concentrated. The concentrate is chromatographed over acid-Washed alumina and eluted with mixtures of ether-petroleum and ether to give 6m-methyl-17achloroethynyl-19-nor-4-androstene-17fl-ol-3-one acetate.

In accordance with the above procedures, but starting with the 60c methyl-17ot-bromoethynyl-19-nor-4-androstene-17/3-ol-3-one in place of the 6a-methyl-17wchloroethynyl-l9-nor 4 androstene-17 3-ol-3-one, there is obtained the 6a-methyl-l7a-bromoethynyl-19-nor-4-androstene-17fl-ol-3-one acetate.

A mixture of 500 mg. of the 6OL-II18thY1-170t-Ch'l0l0- ethynyl-19-nor-4-androstene-17,8-ol-3-one, 10 ml. of dimethyl-formamide, 20 ml. of methyl iodide, and 1.5 gms. of silver oxide is stirred at room temperature for 4 days. An additional /2 gm. of silver oxide is added at the end of each day. One hundred ml. of chloroform is then added to the reaction mixture which is then stirred for one hour, filtered, and the filtrate is evaporated to dryness. The residual oil is chromato-graphed with acid-washed alumina and eluted with mixtures of ether and petroleum ether to give 600 methyl-Not-chloroethynyl-19-nor-4-androstene- 17 3-methoxy-3-one.

-In accordance with the above precedures, but starting with the 60: methyl-17a-bromoethynyl-19-nor 4 androstene-l7 8-ol-3-one in place of the 6a-methyl-17achloroethynyl-19-nor-4-androstene-17,6-01- 3 one, there is obtained the oat-methyl-17abromoethynyl-19-nor-4-androstene-17B-methoxy-3-one.

Example 33 To a solution of 200 mg. of the 17a-chloroethynyl-19- nor-3,5-androstadiene-3,17fi-diol diacetate (Example 17) dissolved in 2 ml. of acetic acid is added, with stirring, 56 ml. of N-chlorosuccinimide and 2 ml. of tetrahydrofuran containing of dry HCl. The reaction is stirred at room temperature for 2V2 hours. Sodium bicarbonate is added. The reaction mixture is then extracted with ether. The ether extracts are ldried and concentrated. The material is chromatographed over acid-washed alumina, and eluted with ether-petroleum ether mixtures to give 6u-ch1oro-Hot-chloroethynyl-19- nor-androstene-17B-ol-3-one acetate.

In accordance with the above procedures, but starting with the 17oz bromoethynyl-19-nor 3,5 androstadiene- 3,17fi-diol diaoetate in place of the Hot-chloroethynyll9-nor-3,5-androstadiene-3,17B-diol diacetate there is obtained the 6a-chloro-17a bromoethynyl 19 nor androstene-17B-ol-3-one acetate.

Example 34 A solution of the 1Wat-chloroethynyl-19-nor-3,5androstadiene-3,17fl-diol diaoetate in aqueous tetra hydrofuran is treated with a slow stream of perohlory-l fluoride for one hour and then allowed to stand at room temperature for an additional 2 hours. The reaction mixture is poured on ice and extracted with chloroform. The chlorofiorm layer is washed with aqueous bicarbonate solution, dried and concentrated in vacuo. The crude material is dissolved in acetic acid saturated with gaseous hydrogen chloride and allowed to stand for one hour at room temperature. The reaction mixture is poured on ice and extracted with chloroform. 'I'he chloroform layer is washed with aqueous bicarbonate solution, dried and concentrated in vacuo. The concentrate is chromatographed on acid-washed alumina to yield the Got-fluoro- 17cc chloroethynyl-19=nor-androstene 17,3 o1 3 one acetate.

In accordance with the above procedures, but starting with the 17a-bromoethynyl-19-nor 3,5 androstadiene- 3-17p2-diol diacetate in place in the 17a-chloroethynyl-19- nor-3,5-androstadiene-3,17fiadiol ldiacetate there is obtained the 6ct-flllOI'O-d'lcHbIOIIlOCthYIlYl 19 nor-4-androstene-17 3*ol3-one acetate.

A mixture of 500 mg. of the '17a-chloroethynyl-19- nor-4-androstene-17fl-ol-3-one (Example 3) 10 ml. of dimethylformamide, 20 ml. of methyl iodide, and 1.5 gm. of silver oxide is stirred at room temperature for 4 days. An additional /2 gm. of silver oxide is added at the end of each day. One hundred ml. of chloroform is then added to the reaction mixture which is then stirred for one 'hour, filtered, and the filtrate is evaporated to dryness. The residual oil is ohromatographed with acid-washed alumina and eluted with mixtures of ether and petroleum ether to give Not-chloroethynyl-19-nor-4- androstene-l7p-methoxy-3-one.

In accordance with the above procedures, but starting with the 17ubromoethynyl-19-nor-4-androstene-17,8-01- 3-one in place of the Net-chloroethynyl-19-nor-4-anldrostrene-17/3+ol-3-one there is obtained the 17a-bromoethynyl-l9 nor-4-androstene-l7/8-methoxy-3-one.

A mixture of 1 g. of 17 tit-chloroethynyl-19-nor-4-androstene-17fi-methoxy-3-one, 10 ml. of acetic anhydride and mg. of p-tol-uenesulfonic acid is heated on the steam bath :for one hour and allowed to stand overnight at room. temperature. It is then poured into ice Water and left to stand at room temperature for one half hour. The reaction mixture is then extracted with ether. The ether extract is washed with Water and aqueous sodium bicarbonate, dried, and concentrated to yield the crude chloroethynyl 19 nor 3,5 andr-ostadiene 17B- methoxy-3-ol acetate.

In accordance with the above procedures, but starting with the 17abromoethynyl-19-nor 4 androstene 17 8- methoxy-3-one in place of the 17u-chloroethynyl19-nor- 4-androstene-17/3-methoxy-3-one, there is obtained the 1701. -bromoethynyl 19 nor 3,5 androstadiene 17,8- methoxy-3-ol acetate.

To a solution of 200 mg. of the Hot-chloroethynyl- 19-nor-3,S-androstadiene-17B-methoxy 3 ol acetate dissolved in 2 ml. of acetic acid is added, with stirring, 56 ml. of N-chlorosuccinimide and 2 ml. of tetrahydrofuran containing 5% of dry I-ICl. The reaction is stirred at room temperature for 2 /2 hours. Sodium bicarbonate is added. The reaction mixture is then extracted with ether. The ether extracts are dried and concentrated. The material is chromatographed over acid-washed alumina, and eluted with ether-petroleum ether mixtures to give Got-chloro-17a-chloroethynyl-19- nor-4-androstene-17fl-methoxy-3-one.

In accordance with the above procedures, but starting with the 17a-bromoethynyl-19-nor-3,5-androstadiene-17fimethoxy-3-ol acetate in place of the 17a-chloroethy-nyl- 19-nor-3,S-androstadiene-17B-methoxy-3-ol acetate there is obtained the 6a-ch loro-17a4bromoethynyl 19 nor-4- androstene-17/3-methoxy-3-one.

Example 35 A solution of the 17OC-ChlOl'O6thYI1Yl-19-I1OT-3,5-3lld1'0- stadiene-17 3-met-hoxy- 3 ol acetate in aqueous tetrahydrofura-n is treated with a slow stream of perchloryl fluoride for one hour and then allowed to stand at room temperature for an additional 2 hours. The reac tion mixture is poured on 'ice and extracted withchloroform. The chloroform layer is washed with aqueous bicarbonate solution, dried and concentrated in vaouo. The crude material is dissolved in'acetic acid saturated with gaseous hydrogen chloride and allowed to stand for one hour at room temperature. The reaction mixture is'poured on ice andex tracted with ohl'oroforml The chloroform layer is Washed with aqueous bicarbonate solution, \dried and concentrated in vacu'ol The concentrate is chromatographed on acid-washed alumina to yield the 6a-fluoro-17a-chloroethynyl-19-nor 4 --androstene-17fl-methoxy-3-one.

In accordance with the above procedures, but starting with the 17a-bromoethyl-l9-nor-3,S-androstadiene-Ufimethxy- 3-ol acetate in place of the 17ot-chloroethynyl-19- nor-3,5-androstadiene-l7fl-methoxy-3-ol acetate, there is obtained the 6a-fiuoro-17a-bnomoethynyl-19-nor-4 andrstene-l7fi-rne thoxy-3mne.

Example 36 A solution of 100 mg. of the 17u-chlor0ethynyl-19-nor- 4-.androstene-17B-ol-3-one and 50 mg. ofLindlar catalyst in cc. of ethyl acetate is treated with hydrogen until one nmle of hydrogen has been absorbed. The mixture is filtered and concentrated :to yield the crude chlorov-inyl compound, Chromatography yields the pure product 21- chloro-19-nor-4-,20 pregnacliene-17fl-ol-3-one.

In accordance with the above procedure, but starting with the l7ot-bromoethynyl-l9-nor-4 androsten-17 8-01-3- one, the 17a-chloroethynyl-19-nor-5 (10)-androstene-17B- ol-3-one, or the 17abromoethynyl-l9-n0r-5(10)-androstene-17fl-ol-3-one, in place of the 17 a-chloro-ethynyl-19- nor-4-andnostene-17(3-ol-3-0ne, there is obtainedthe 2-1- bromo-l9-nor-4,20-pregnadiene-17,8-ol-Sone, 21-chloro- 19-nor-5 (10),20-pregnadiene-17fi-ol-3-one, or 21-bromo l9-nor-5 (10),20-pregnadieneel7fi-ol-3-one respectively.

- Example 37 A suspension of platinum oxide in 10 cc. of ethanol '32 is reduced and 100 mg. of 17a=chloroe thynyl-l9-nor-4- androstene-Ufl-ol-B-one is added. Reduction proceeds until two moles of hydrogen have been absorbed. The solution is filtered, concentrated and chromatographed on alumina 'to'yield the 21-chloro-19-nor-4-pregnene-17-ol- 3-one.

In accordance with the above procedure, but starting with the 17a-brornoethynyl-l9 nor-4 androstene-1713-01-3- one, the 17a-chloroethynyl-19-nor-5 10') -andr'ostene-17pol-3-one or the 17a hromoethynyl-19-nor-5 10) -androstene-l7fi ol-3-one in place of the i17a-chloroethynyl-19- nor-4*androstene-l7/3-ol-3 oiie, there is obtained the 21- bromo-l9-nor 4-pregnene17 3-ol-3-one, the 21'-chloro-19- nor-5 (lOypregnene- 1 -ol-3-one', or the 21-bromo-19-nor- 5 10) -pregnene-17{3ol-3=one respectively.

We claim:

' 1. Process for the preparation of 17u-ethynyl-5-andr0- stem-3,8,17fl-diol-bis tetrahydropyranyl ether which comprises reacting 17a-ethynyl-5-androstene-35,17(3-diol with dihydropyran in the presence of an acidic reagent.

2. Process for the preparation of 17u-ha1oethynyl-5- andrOStene-Bfl,17fl-diol-bis tetrahydnopyranyl ether which comprises halogenating 17a-ethynyl-5-androstene-3fi,17B- diol-bis-tetrahydropyranyl ether.

3. Process for the preparation of l7a-haloethynyl-5- androstene-3B,17B-dio1which comprises reacting 17u-haloethynyl 5 androstene 35,173 diol bis tetrahydropyrany-l ether with concentrated HQI.

4. Process for the preparation of 17u-haloethynyl-4-androstene-lTfl-ol-S-dne which comprises reacting 17a-haloethynyl-5-androstene-3fl-diol with cyclohexanone in the presence :of aluminium isopropoxide in an organic solvent.

5. Process for the preparation of 17a-haloethynyl-19- nor-4-androstene-17fi-ol-3one which comprises reacting a mixture of A and A -17u-ha11oethynyl-3-ethylenedioxy- 19-nor-androstene-1713*ol-tetrahydropyrany1 ethers with concentrated HCl. 7

No references cited. 

1. PROCESS FOR THE PREPARATION OF 17A-ETHYNYL-5-ANDROPRISES REACTING 17A-ETHYNYL-5-ANDROSTENE-3B,17B-DIOL WITH DIHYDROPYRAN IN THE PRESENCE OFF AN ACIDIC REAGENT.
 2. PROCESS FOR PREPARATION OF 17A-HALOETHYNYL-5STENE-3B,17B-DIOL-BIS-TETRAHYDROPYRANYL ETHER WHICH COMANDROSTENE-3B,17B-DIOL-BIS-TETRAHDROPYRANYL ETHER WHICH COMPRISES HALOGENATING 17A-ETHYNYL-5-ANDROSTENE-3B,17BDIOL-BIS-TETRAHYDROPRANYL ETHER. 